Five years after the collapse of the Fundão Dam: lessons from temporal monitoring of chemistry and acute toxicity.

In November 2015, the Fundão Dam break released millions of tons of metal-rich tailings into the Doce River Basin (DRB), causing catastrophic damage and potential ecological effects that reached the Atlantic Ocean. This study aimed to evaluate the geochemistry and toxicity of water and sediments collected in the DRB from 2015 to 2019 and to determine the spatial and temporal trends. Water and sediment samples were analyzed for metals and As by inductively coupled plasma optical emission spectrometry (ICP-OES), and acute toxicity for Daphnia similis or D. magna. Results were explored using geochemical indices and correlation analyzes. Overall, higher concentrations of metals and As in water and sediments were observed immediately after dam breakage, but the levels exhibited a decreasing trend over time, although the levels of some elements such as As and Mn remained high in the upper DRB. The geochemical indices indicated mostly low to moderate contamination, and the enrichment factor (EF) demonstrated a higher enrichment of Mn in the upper DRB. Acute toxicity to water fleas (D. similis and D. magna) was occasionally observed in waters and sediments, but the reference samples were toxic, and the short-term effects were not correlated with metals and As. Overall, the results showed limited bioavailability of metals and As and a decreasing trend in their concentrations, indicating an ongoing recovery process in DRB. These results are important to decision-making regarding the disaster and actions for environmental restoration.

Linking MS1 and MS2 signals in positive and negative modes of LC-HRMS in untargeted metabolomics using the ROIMCR approach.

Data-independent acquisition (DIA) mode in liquid chromatography (LC) high-resolution mass spectrometry (HRMS) has emerged as a powerful strategy in untargeted metabolomics for detecting a broad range of metabolites. However, the use of this approach also represents a challenge in the analysis of the large datasets generated. The regions of interest (ROI) multivariate curve resolution (MCR) approach can help in the identification and characterization of unknown metabolites in their mixtures by linking their MS1 and MS2 DIA spectral signals. In this study, it is proposed for the first time the analysis of MS1 and MS2 DIA signals in positive and negative electrospray ionization modes simultaneously to increase the coverage of possible metabolites present in biological systems. In this work, this approach has been tested for the detection and identification of the amino acids present in a standard mixture solution and in fish embryo samples. The ROIMCR analysis allowed for the identification of all amino acids present in the analyzed mixtures in both positive and negative modes. The methodology allowed for the direct linking and correspondence between the MS signals in their different acquisition modes. Overall, this approach confirmed the advantages and possibilities of performing the proposed ROIMCR simultaneous analysis of mass spectrometry signals in their differing acquisition modes in untargeted metabolomics studies.

Dredging impacts on the toxicity and development of sediment quality values in a semi-arid region (Ceará state, NE Brazil).

Sediment dredging impacts coastal environments by promoting the resuspension of fine particles and remobilization of contaminants that may trigger toxic effects. In this study, we evaluated the sediment quality in harbor areas of Mucuripe bay, a semi-arid ecosystem located in Ceará state (Brazil), which is subject to dredging activities. A sampling survey was conducted right after dredging operations and data compared to another survey performed prior dredging. Sediments were analyzed for fine particles, organic carbon, nutrients, metals, hydrocarbons, and tributyltin (TBT). Toxicity of whole-sediment and liquid phase exposures were also determined. The concentrations of Cd, Cr, Cu, and Zn decreased after dredging, which was confirmed by the geoaccumulation index. Levels of TBT dropped while phosphorus, aliphatic and polycyclic aromatic hydrocarbons increased. Toxic effects persisted, indicating a post-dredging recontamination combined with other sources such as urban runoff, wastewater discharges, harbor activities, and antifouling particles. Data from Mucuripe and Pecém harbors were compiled and site-specific sediment quality values (SQVs) were developed by using multivariate methods. The threshold values proposed by our study were lower and more effective to predict toxicity compared to international guidelines, indicating levels of contamination for this tropical region in which toxic effects may occur. Considering the large geographic area with different sediment characteristics of the Brazilian coast, this study represents a significant contribution to sediment toxicity assessment of dredging activities in semi-arid environments.

Levels of hydrocarbons and toxicity of water-soluble fractions of maritime fuels on neotropical invertebrates.

Accidents involving fuels and oil spills are among the main sources of hydrocarbons to the marine ecosystems and often damage the biota. Diesel and bunker oil are two examples of fuels with broad application that release hydrocarbons to the aquatic environment and little is known about their toxicity on tropical organisms. This study aimed to assess the toxicity of the water-soluble fraction (WSF) of diesel and bunker oils to neotropical marine invertebrates. Commercial fuels were purchased for WSF extraction, analyzed for total petroleum hydrocarbons (TPH), and polycyclic aromatic hydrocarbons (PAHs), and acute and chronic toxicity determined. The WSF analyzed contained varied levels of TPH and PAHs mixtures, especially low molecular weight PAHs; bunker WSF presented higher amounts of TPH and PAHs. Both WSFs tested produced significant mortality of the brine shrimp Artemia salina, affected the reproduction rate of the copepod Nitokra sp, and impaired the embryo-larval development of the mussel Perna perna and of the sea urchin Lytechinus variegatus. In general WSF from diesel was more toxic to the organisms tested, but bunker WSF was more toxic to embryos of L. variegatus. Toxicity started from concentrations of 3% WSF, which can be environmentally relevant after an oil spill, indicating that marine biota may be adversely affected in short term.

Sub-lethal Responses of the Polychaete Armandia agilis in Whole-sediment Toxicity Testing.

The present study assessed biochemical responses as sublethal endpoints in the polychaete Armandia agilis exposed to contaminated sediments to in order to assess its potential use as a test organism. Sediment samples from several locations at a dredging site were obtained and used in whole-sediment exposures. Samples were tested with A. agilis to determine the 10-day toxicity of the 100% sample and the enzymatic activity of catalase (CAT), glutathione-S-transferase (GST) and acetylcholinesterase (AChE) biochemical measurements made in whole-body homogenates of a subset of the surviving organisms. Biochemical responses reported in A. agilis were not statistically different from the reference site sediment, however, the integrated analysis demonstrated that contaminants bound to sediment samples influenced the sublethal effects.

PAH residues and toxicity levels two years after an extensive oil spill on the northeast Brazilian coast.

The most extensive oil spill ever recorded in tropical oceans occurred between August 2019 and March 2020, affecting approximately 3000 km of the Brazilian coast. This study assessed the chemical contamination and toxicity of sediments collected from affected reef areas during two sampling surveys conducted 17 and 24 months after the peak of oil slick inputs. Our results indicated that neither PAH levels nor measured toxicity showed a significant contribution from the spilled oil, with concentrations and biological effects indistinguishable from those in unaffected areas. Similarly, no differences were observed between seasons. Furthermore, there was no discernible relationship between sediment toxicity results and the measured PAH concentrations. Therefore, while biological responses indicated toxicity in most assessed areas, these responses are likely related to other local sources. This evidence suggests a natural oil attenuation process contributing to local environmental recovery. Nonetheless, further investigation is needed for other areas affected by oil spills.

Unraveling sources of hydrocarbons in subtropical estuaries with distinct degrees of protection in the Southwestern Atlantic Ocean, Brazil.

Sedimentary aliphatic hydrocarbons and polycyclic aromatic hydrocarbons (PAHs) were studied in subtropical estuaries with distinct degrees of legal protection, located in the Southwestern Atlantic Ocean, São Paulo State, Brazil. A multivariate approach was adopted, using the Hierarchical cluster analysis followed by the Principal Matrix Factorization analysis to support the hydrocarbons sources findings using diagnostic ratios. In general, the sites with the highest values of hydrocarbons were in the vicinity of well-urbanized cities, where sewage discharge, harbor and industrial activities take place. Pyrolitic PAHs were the predominant source of PAHs in the sites. The region can be considered not highly contaminated by hydrocarbons; however, specific sites under local anthropogenic impact from sewage and/or urban drainage, presented relatively high hydrocarbons levels. These findings highlight the importance of sources identification as reliable approach to be included in the management plan of protected areas under the inputs of several vectors of contamination.

Interactive effects of microplastics and benzo[a]pyrene on two species of marine invertebrates.

This study aimed to evaluate B[a]P and low-density polyethylene microplastics (MPs) toxicty, alone and in mixture (0.03 to 30 µg L-1 of B[a]P; and 5, 50 and 500 mg L-1 for MPs). Five mg L-1 of MPs is considerably higher than commonly reported environmental concentrations, although it has been reported for marine environments. Individual (sea urchin embryo-larval development and mortality of mysids) and sub-individual responses (LPO and DNA damage in mysids) were assessed. The toxicity increased as the B[a]P concentration increased, and microplastics alone did not cause toxicity. B[a]P toxicity was not modified by the lowest concentration of MPs (5 mg L-1), but at higher MPs concentrations (50 and 500 mg L-1), the effects of B[a]P on sea urchin development and in biomarkers in mysids were diminished. Microplastics interacted with B[a]P in seawater, reducing its toxicity, probably due to adsorption of B[a]P to the surface of microplastics.

Using large amounts of firefighting foams releases per- and polyfluoroalkyl substances (PFAS) into estuarine environments: A baseline study in Latin America.

We analyzed per- and polyfluoroalkyl substances (PFAS) in aqueous film-forming foams (AFFFs) used to extinguish a major fire in a petrochemical terminal from the Port of Santos (Brazil). Eight AFFFs from seven known commercial brands and one unknown sample (AFFF-1 to AFFF-8) were evaluated. 17 PFAS were identified and quantified using high performance liquid chromatography (LC/MS). The concentrations of Σ17 PFAS in the AFFFs ranged from 500 to 9000 ng/g, with prevalence of short chain PFAS (~85 %), followed by long chain PFAS. Perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA), included in the global treaty of the Stockholm Convention, were also detected. We estimated that at least 635.96 g of PFAS were introduced in the estuary, representing a massive input of these substances. This investigation reports the PFAS composition of AFFFs used in firefighting in the GRULAC Region (Group of Latin American and Caribbean countries).

From molecular endpoints to modeling longer-term effects in fish embryos exposed to the elutriate from Doce River.

Sediments represent a major sink and also a main source of contaminants to aquatic environments. An environmental disaster from a mining dam breakage in 2015 in South-East Brazil re-suspended complex mixtures of chemicals deposited in the sediment, spreading contaminants along the Doce River Basin (DRB) major river course. While high levels of contaminants in sediment were well described, toxicological effects in aquatic organisms were poorly investigated. Thus, the effects of these potentially toxic chemicals were assessed in the present study through different endpoints (biochemical to populational levels) in fish embryos of the South-American silver catfish exposed to elutriates from different sites of the DRB. Despite no significant mortality observed, our results showed that exposure to the elutriates, especially those from the closest site to the dam collapse, caused higher deformities rates and DNA damage in the fish embryos than in the control group. Multivariate analysis showed that these sublethal effects may be related to the high levels of metals introduced by mining activities, compromising long-term survival and reproduction success. In addition, it was possible to observe the influence of other sources of pollutants along the river. According to our data, the mathematical model simulated a significant impact on the population density at longer-term exposure, for the sites that showed the most prominent toxicity responses. The fish embryo toxicity test proved to be an effective assay to assess the ecotoxicological effects of the pollutants from a major river contaminated by a mining dam collapse and showed that the survival rate per se was not a suitable endpoint to assess the toxicity of the pollutants. As a consequence, we contributed to shed a light on a potential underestimated impact of pollutants in sediments of the DRB on the native organisms at distinct biological levels of organizations.

Univariate or multivariate approaches for histopathological biomarkers in the context of environmental quality assessments?

Although the simplification of multivariate histopathological data into univariate indices can be useful for the assessment of environmental quality, this implies a great loss of information. The objective of the present study was to evaluate the effectiveness, in the context of environmental quality assessment, of an approach that integrates individual histopathological responses in a discriminated manner with the results of contaminants by means of multivariate analyses. This analysis was compared to the diagnosis of environmental quality provided by the use of the univariate Bernet histopathological index. Contaminant loads (sediments and fish) and the liver histopathology of Cathorops spixii were integrated through multivariate analysis. Integrated individual histopathological responses allowed classifying environmental quality from more to less impacted sites, while the univariate index showed some inconsistencies with chemical loads and allowed identifying only the most impacted site.

Integrated analysis of fish intestine biomarkers: Complementary tools for pollution assessment.

The gastrointestinal tract and its enteric nervous system are the first routes of food and xenobiotics uptake. Considering the importance of this organ, this study evaluated intestinal biomarkers of Sphoeroides testudineus integrating the data to generate tools for pollution assessment. The fish were collected in three sites of São Paulo Coast and their intestines were analyzed for biochemical, histology, and neuronal density and morphometry biomarkers. To evaluate the differences among the data, a PERMANOVA was applied, followed by a FA/PCA. The PERMANOVA indicated differences (P < 0.001) between the regions (RA, A1, and A2). Four factors were extracted from the FA/PCA (62% cumulative), showing that the animals from A2 presented severe alterations, mainly in intestinal morphometry and neuronal density. A1 alterations refer mainly to the increase of neuronal metabolism. Our results also evidence a gradient of environmental quality related to the protection level (AR > A1 > A2).

Non-destructive biomarkers can reveal effects of the association of microplastics and pharmaceuticals or personal care products.

Methods to assess the effects of contaminants on marine organisms typically involve euthanasia to obtain samples, but less invasive techniques may be more appropriate for working with threatened species. In this study, were assessed the biological responses of crabs exposed to microplastics and contaminants of emerging concern. Biochemical and cellular effects (lipid peroxidation, DNA damage, cholinesterase activity, and lysosomal membrane stability) in hemolymph were analyzed in a kinetic study, at 3 and 7 days, in U. cordatus exposed to microplastics spiked with Triclosan (TCS) or 17α-Ethynylestradiol (EE2). The results showed that the contaminants were produced toxic effects in the crabs exposed either to the microplastics alone (oxidative stress, genotoxicity, and neurotoxicity), or to microplastics with TCS or EE2 adsorbed (neurotoxic and cytotoxic). The present study showed the responsiveness of non-lethal analyzes to understanding the biological effects of combined exposure to microplastics and chemical pollution.

Fire Suppression Agents Combined with Gasoline in Aquatic Ecosystems: A Mixture Approach.

Fire suppression agents are recommended for extinguishing fires by flammable liquids and frequently end in water bodies, combined with the fuels. There is a lack of toxicity information on these commercial formulations and the effects of mixtures of fire suppression agents and fuels. The aim of the present study was to evaluate the toxic effects of different fire suppression agents, the gasoline water-soluble fraction (GWSF), and mixtures of each fire suppression agent and GWSF. Individual tests were performed with Daphnia similis and Artemia sp.; the most toxic fire suppression agents to D. similis and Artemia sp. were F-500®, Cold Fire®, Agefoam®, and Kidde Sintex® 1%; the GWSF was the least toxic. The concentration addition model was used to predict the mixture effects and evaluate synergism/antagonism, dose ratio dependence, and dose level dependence. Cold Fire with GWSF showed dose level deviation to D. similis, marked mainly by synergism; for Artemia sp., the dose ratio pattern was predicted, with a synergistic response mainly by Cold Fire. Agefoam and GWSF behaved additively for D. similis and dose ratio for Artemia sp., with synergism being caused by Agefoam. Kidde Sintex 1% with GWSF were dose ratio for both organisms, with Kidde Sintex 1% being responsible for synergism. Our results show that some mixtures of fire suppression agents and GWSF may cause toxicity to aquatic organisms, posing risk in a real environmental scenario, such as a major fire combat. Environ Toxicol Chem 2021;40:767-779. © 2020 SETAC.

Sediment contamination and toxicity of Marine Protected Areas in southeastern Brazilian coast: A baseline for management planning.

Recent studies have shown that the effectiveness of establishing Marine Protected Areas (MPAs) is threatened by toxic effects of contaminants. São Paulo is the most economically developed state of Brazil, and its coast is protected by a system of MPAs, including the North Shore Marine Protected Area (NSMPA). The present study provides a first assessment of sediment quality in NSMPA and two reference sites. Concentrations of arsenic (As), metals (Al, Fe, Cd, Cr, Cu, Ni, Pb, and Zn) and polycyclic aromatic hydrocarbons (PAHs) and its sources were determined. Comparisons with sediment quality guidelines indicated low contamination, as confirmed by the Geoaccumulation Index, which classified the results as background levels. Whole-sediment exposures of the amphipod Tiburonella viscana were performed to estimate possible acute effects, but samples exhibited no toxicity. The results presented in this study represent a baseline for further monitoring programs of MPAs, supporting conservation actions of management plans.

Adaptative responses of myenteric neurons of Sphoeroides testudineus to environmental pollution.

Contamination in estuarine regions affects the local biota damaging the ecosystems and reaching humans. The gastrointestinal tract is a dynamic environment capable of obtaining nutrients and energy from food while it protects the host against harmful toxins and pathogens from the external environment. These functions are modulated by the enteric nervous system and changes in its structure can result in gastrointestinal disorders. The objective of this study was to evaluate if the environmental contaminants have effects on the myenteric neuronal plasticity of pufferfish Sphoeroides testudineus. Animals were collected in Barra do Una River, located at Jureia-Itatins Mosaic of Protected Areas (reference area - RA) and in the Santos Estuarine System (impacted area - IA). Morpho-quantitative analyses of the general and metabolically active myenteric neuronal populations of the proximal and distal intestine were made. Disarrangement was observed in the general organization of the myenteric plexus, with an expressive reduction of the neuronal groups (nodes) in the animals of IA. The vulnerability of the myenteric plexus was evidenced by a decrease in density and cellular profile of the general neuronal population, followed by an increase of the metabolism of the remaining neurons, which in turn was verified by a growth of the area of the cellular and nuclear profiles of the metabolically active neuronal population. Through these analyses, we concluded that animals inhabiting polluted regions present alterations in the myenteric neuronal plasticity, as a way of maintaining the functions of the gastrointestinal tract.

Toxicity of three emerging contaminants to non-target marine organisms.

Coastal areas are continually impacted by anthropic activities because they shelter large urban conglomerates. Urban effluents directly or indirectly end up reaching the marine environment, releasing a large number of pollutants which include the so-called contaminants of emerging concern (CECs), since the conventional treatment plants are not effective in removing these compounds from the effluents. These substances include hormones, pharmaceuticals and personal care products, nanoparticles, biocides, among others. The aim of this study was to evaluate the toxicity of the 17α-ethinylestradiol (EE2), acetylsalicylic acid (ASA), and bisphenol-A (BPA) to two marine crustaceans and one echinoderm, evaluating the following parameters: survival (Artemia sp. and Mysidopsis juniae), embryo-larval development (Echinometra lucunter). The LC50 values calculated in the acute toxicity tests showed that the compounds were more toxic to M. juniae than to the Artemia sp. Among the three contaminants, EE2 was the most toxic (LC50-48h = 18.4 ± 2.7 mg L-1 to Artemia sp.; LC50-96h = 0.36 ± 0.07 mg L-1 to M. juniae). The three tested compounds affected significantly the embryonic development of the sea urchin in all tested concentrations, including ecologically relevant concentrations, indicating the potential risk that these contaminants may present to the marine biota.

Combined effects of temperature and copper on oxygen consumption and antioxidant responses in the mudflat fiddler crab Minuca rapax (Brachyura, Ocypodidae).

This study investigates the combined effects of waterborne copper exposure and acute temperature change on oxygen consumption and the oxidative stress biomarkers, glutathione S-transferase (GST) and glutathione peroxidase (GPx), in the gills and hepatopancreas of the fiddler crab Minuca rapax. Crabs held at 25 °C were acclimated to 0 (control), 50, 250 or 500 µg Cu L-1 for 21 days, and were then subjected to 15, 25 and 35 °C for 24 h. Aerial oxygen consumption rates of crabs in copper free media increased with increasing temperature from 15 to 35 °C, Q10 values reaching ≈3. Crabs exposed to increasing copper concentrations exhibited variable responses, Q10 values falling to ≈1.5. Copper had no effect on oxygen consumption at 25 °C. However, at 35 °C, rates decreased in a clear concentration-response manner in the copper exposed crabs, revealing impaired aerobic capability. At 15 °C, oxygen consumption rates increased with copper concentration, except for a decrease at 500 µg Cu L-1. Gill GST activity was ≈2-fold that of the hepatopancreas, while hepatopancreas GPx activity was 3-fold that of the gills. Gill GST activities were reduced by copper exposure only at 25 °C while hepatopancreas GST activities were altered by copper at all temperatures. Hepatopancreas GST and GPx activities increased in crabs exposed to copper at 35 °C, revealing oxidative stress induction. Hepatopancreas GST and GPx activities were reduced in copper exposed crabs at 15 °C, suggesting a diminished capability to mitigate the effects of copper exposure at low temperature. These findings reveal that copper exposure increases oxygen consumption at low temperatures but decreases consumption at high temperature. Hepatopancreas GPx activities decreased at low temperature and increased at high temperature. These novel findings demonstrate that the interaction between copper exposure and temperature should be considered when evaluating biomarker activities in semi-terrestrial crabs.

Multiple lines of evidence of sediment quality in an urban Marine Protected Area (Xixová-Japuí State Park, SP, Brazil).

Marine Protected Areas (MPAs) aim to protect habitats, biodiversity, and ecological processes as a conservation tool. These areas have been affected by contamination, which threats the biodiversity and ecological functioning. In this study, we evaluated the sediment quality of Xixová-Japuí State Park (XJSP), an MPA located in an urbanized Bay (Santos, Southeast Brazil) by integrating multiple lines-of-evidence. Six sites were selected within the XJSP and analyzed for sediment chemistry, toxicity, and benthic community descriptors using Sediment Quality Triad approach (SQT). Whole-sediment Toxicity Identification Evaluation (TIE) was employed as a complementary line of evidence to confirm the presence of domestic effluent discharges as a potential stressor. The SQT showed that sediments collected within XJSP are impacted by contaminants, exhibiting chronic toxicity and changes in benthic community. TIE results indicated that trace metals, organic contaminants, and ammonia contributed to the observed effects. Our results also indicate a lack of effectiveness of MPA in protecting the biodiversity due to the contamination sources, which requires efforts to pollution control in order to ensure the environmental conservation and management plan goals.

Detoxification, oxidative stress, and cytogenotoxicity of crack cocaine in the brown mussel Perna perna.

The presence of cocaine and its metabolites and by-products has been identified in different aquatic matrices, making crack cocaine the target of recent studies. The aim of this study was to evaluate the sublethal effects of crack on the brown mussel Perna perna. Mussels were exposed to three concentrations of crack cocaine (0.5, 5.0, and 50.0 µg L-1) for 168 h. Gills, digestive glands, and hemolymph were extracted and analyzed after three different exposure times using a suite of biomarkers (EROD, DBF, GST, GPX, LPO, DNA damage, ChE, and lysosomal membrane stability [LMS]). After 48 and 96 h of exposure, EROD, DBF, GST, GPX activities and DNA strand breaks in the gills increased significantly after 48 and 96 h of exposure. Alterations in LMS were also observed in the mussels exposed to all crack concentrations after 96 and 168 h. Our results demonstrated that crack cocaine is metabolized by CYP-like and GST activities in the gills. GPX was not able to prevent primary genetic damage, and cytotoxic effects in the hemocytes were also observed in a dose- and time-dependent response. Our study shows that the introduction of illicit drugs into coastal ecosystems must be considered a threat to marine organisms.