A multi-biomarker approach to verify chronic effects on Eisenia andrei earthworms exposed to tailings from one of the world's largest mining disasters.

Mining is of great relevance to the global economy, but its activities are challenging due to socio-environmental impacts. In January 2019, an iron ore tailings dam collapsed in Brumadinho (Minas Gerais, Brazil) releasing 12 × 106 m3 of tailings, causing human losses and devastation around 3.13 × 106 m2 of a watershed. In this context, the present study aimed to investigate the potential toxic effects of tailings from the collapsed dam using earthworms Eisenia andrei as a model organism for terrestrial environments. An extensive set of tests was performed, including behavioral (avoidance), acute (mortality and biomass) and chronic tests, such as biomass, reproduction and cytotoxicity (viability and cell density and change in coelomocyte pattern). The physical-chemical characterization revealed a higher density of the tailings in relation to the control soil, which can result in physical changes, such as soil compaction and surface sealing. Aluminum, Ca, Fe, Hg, Mg, Mn, K, Na and P registered higher concentrations in the tailings compared to the control soil, while Total Nitrogen, Total Organic Carbon and Organic Matter were higher in the natural soil. Based on the avoidance test, an EC50 of 27.18 ± 2.83% was estimated. No lethality was observed in the acute exposure, nor variations in biomass in the acute and chronic assays. However, there was a tendency to reduce the number of juveniles in relation to cocoons in the proportions of 3125; 12.5 and 25%. Significant changes in viability, cell density and pattern of amebocytes and eleocytes were observed up to the 35th day of exposure. A multi-biomarker approach (Integrated Biological Response version 2) indicated concentration-dependent effects and attenuation of cellular changes over time. These are the first results of chronic effects on earthworms exposed to tailings from the B1 dam. Despite being conclusive, we highlight the possible heterogeneity of the tailings and the necessary care in extrapolating the results.

Antioxidant system alterations, oxidative, and genotoxic effects in Danio rerio (zebrafish) exposed to leachate from a dumpsite.

Water body contamination by leachate originated from dumpsites is a concern for municipal solid waste (MSW) management. In this context, this study aimed to evaluate antioxidant system alterations and oxidative and genotoxic effects in Danio rerio (zebrafish) exposed to leachate from a closed dumpsite. Groups comprising 50 fish were exposed (96 h) to different leachate concentrations (5, 15, 30, and 50%) to evaluate effects on liver and brain superoxide dismutase (SOD), catalase (CAT), and glutathione-S-transferase (GST) activities and reduced glutathione (GSH) and metallothionein (MT) concentrations, as well as malondialdehyde (MDA) and protein carbonylation (PTC) levels. Blood genotoxicity was evaluated by the comet assay. The investigated dumpsite leachate pond presented high chloride concentrations (Cl-; 2288.4 ± 69.5 mg L-1) and high electrical conductivity (EC; 8434.0 mS cm-1), indicating the presence of leachate. Concerning Danio rerio exposure, higher SOD (37%), CAT (67%), and GST (39%) activities and higher GSH (57%) concentrations were observed in liver following exposure to 50% leachate, while decreased brain GST (42%) activities and GSH (90%) levels were observed at the same leachate concentration. A significant increase in the olive tail moment (OTM; 280%) indicative of genotoxicity in blood was observed. A principal component analysis indicated that increased enzymatic activities and high levels of both GSH and MT were not sufficient to prevent the accumulation of reactive oxygen species, resulting in PTC and genotoxicity. Therefore, leachate exposure causes sublethal Danio rerio effects, altering the antioxidant system, increasing ROS production, and leading to PTC and genotoxicity. The findings demonstrate the need to further develop sublethal level assessments in zebrafish using leachate from different sources to subsidize risk assessments regarding MSW management.

A comprehensive overview on solid waste leachate effects on terrestrial organisms.

Leachate is a highly complex waste with high toxicological potential that poses a significant threat to the terrestrial environment. Determining leachate physicochemical parameters and identifying xenobiotics alone is, however, not enough to determine the real environmental impacts. In this context, the use of terrestrial model organisms has been highlighted as a tool in ecotoxicological leachate assessments and as a guiding principle in risk assessments. In this context, this review aimed to present the most current state of knowledge concerning leachate toxicity and the bioassays employed in this evaluation concerning terrestrial plants and animals. To this end, a literature search on leachate effects on terrestrial organisms was carried out using ten search terms, in 32 different combinations, at the Web of Science and Scopus databases. A total of 74 eligible articles were selected. The retrieved studies analyzed 42 different plant and animal species and employed nine endpoints, namely phytotoxicity, genotoxicity, bioaccumulation, antioxidant system, cytotoxicity, reproduction, physiological changes, behavior and lethality. A frequent association of toxic leachate effects with metals was observed, mainly Pb, Cd, Cr, Mg, Zn and Cr, which can cause antioxidant system alterations and cyto- and genotoxicity. These elements have also been associated to reproductive effects in earthworms and mice. Specifically concerning plants, most of the retrieved studies employed Allium cepa in toxicity assays, reporting phytotoxic effects frequently associated to metals and soil parameter changes. Animal studies, on the other hand, mostly employed mice and evaluated genotoxicity and antioxidant system effects. Even with the description of toxic leachate effects in both plants and animals, a lack of knowledge is still noted concerning reproductive, physiological, cytotoxic, and behavioral effects in terrestrial species. We, thus, suggest that further studies be carried out on other animals, advancing our understanding on potential environmental leachate effects, also allowing for human health risk assessments.

Acute toxicity of single and combined rare earth element exposures towards Daphnia similis.

The increasing use of Rare Earth Elements (REE) in emerging technologies, medicine and agriculture has led to chronic aquatic compartment contamination. In this context, this aimed to evaluate the acute toxic effects of lanthanum (La), neodymium (Nd) and samarium (Sm), as both single and binary and ternary mixtures on the survival of the microcrustacean Daphnia similis. A metal solution medium with (MS) and without EDTA and cyanocobalamin (MSq) as chelators was employed as the assay dilution water to assess REE bioavailability effects. In the single exposure experiments, toxicity in the MS medium decreased following the order La > Sm > Nd, while the opposite was noted for the MSq medium, which was also more toxic than the MS medium. The highest MS toxicity was observed for the binary Nd + La (1:1) mixture (EC50 48 h of 11.57 ± 1.22 mg.L-1) and the lowest, in the ternary Sm + La + Nd (2:2:1) mixture (EC50 48 h 41.48 ± 1.40 mg.L-1). The highest toxicity in the MSq medium was observed in the single assays and in the binary Sm + Nd (1:1) mixture (EC50 48 h 10.60 ± 1.57 mg.L-1), and the lowest, in the ternary Sm + La + Nd (1:2:2) mixture (EC50 48 h 36.76 ± 1.54 mg.L-1). Concerning the MS medium, 75 % of interactions were additive, 19 % antagonistic, and 6 % synergistic. In the MSq medium, 56 % of interactions were synergistic and 44 % additive. The higher toxicity observed in the MSq medium indicates that the absence of chelators can increase the concentrations of more toxic free ions, suggesting that the MS medium should be avoided in REE assays. Additive interactions were observed in greater or equivalent amounts in both media and were independent of elemental mixture ratios. These findings improve the understanding of environmental REE effects, contributing to the establishment of future guidelines and ecological risk calculations.

Zebrafish (Danio rerio) as a model to assess the effects of cocaine as a drug of abuse and its environmental implications.

Cocaine (COC) use concerns are on the increase for both authorities and civil society. Despite this, it is important to investigate COC effects or those of its main metabolite, belzoylecgonine (BE), in consolidated aquatic model organisms, such as the zebrafish (Danio rerio). This (mini) review consists in an assessment regarding toxicological studies carried out employing zebrafish (embryos, larvae or adults) exposed to COC and/or BE indexed at the SCOPUS and Web of Science databases. Ten different endpoints were analyzed in both embryos and larvae, whereas only four were analyzed in adults. Of the 23 studies, only five investigated COC and/or BE effects following an environmental approach when exposing zebrafish, while most (18 studies) analyzed COC effects under a drug of abuse approach. Cocaine exposure was noted as altering the expression of several genes, such as those linked to COC transport proteins, dopamine receptors, SP substance production, the tachykinin system, and the tyrosine hydroxylase enzyme. BE exposure resulted in more oxidative and proteomic effects than COC in embryos. Cocaine abstinence resulted in hyperactivity associated with stereotypy in adult fish, in addition to reduced responses to visual stimuli to red light and neuronal development pattern alterations. Cocaine was noted as accumulating in zebrafish eyes, possibly due to melanin binding, and causing dose-response cardiac effects in both embryos and adults. Despite the different effects addressed by our survey, we emphasize the lack of COC and BE exposure assessments in zebrafish employing an environmental point of view.

A comprehensive assessment of leachate contamination at a non-operational open dumpsite: mycoflora screening, metal soil pollution indices, and ecotoxicological risks.

The final disposal of municipal solid waste (MSW) in dumpsites is still a reality worldwide, especially in low- and middle-income countries, leading to leachate-contaminated zones. Therefore, the aim of this study was to carry out soil and leachate physicochemical, microbiological, and toxicological characterizations from a non-operational dumpsite. The L-01 pond samples presented the highest physicochemical parameters, especially chloride (Cl; 4101 ± 44.8 mg L-1), electrical conductivity (EC; 10,452 ± 0.1 mS cm-1), and chemical oxygen demand (COD; 760 ± 6.6 mg L-1) indicating the presence of leachate, explained by its close proximity to the landfill cell. Pond L-03 presented higher parameters compared to pond L-02, except for N-ammoniacal and phosphorus levels, explained by the local geological configuration, configured as a slope from the landfill cell towards L-03. Seven filamentous and/or yeast fungi genera were identified, including the opportunistic pathogenic fungi Candida krusei (4 CFU) in an outcrop sample. Regarding soil samples, Br, Se, and I were present at high concentrations leading to high soil contamination (CF ≤ 6). Pond L-02 presented the highest CF for Br (18.14 ± 18.41 mg kg-1) and I (10.63 ± 3.66 mg kg-1), while pond L-03 presented the highest CF for Se (7.60 ± 1.33 mg kg-1). The most severe lethal effect for Artemia salina was observed for L-03 samples (LC50: 79.91%), while only samples from L-01 were toxic to Danio rerio (LC50: 32.99%). The highest lethality for Eisenia andrei was observed for L-02 samples (LC50: 50.30%). The applied risk characterization indicates high risk of all proposed scenarios for both aquatic (RQ 375-909) and terrestrial environments (RQ > 1.4 × 105). These findings indicate that the investigated dumpsite is contaminated by both leachate and metals, high risks to living organisms and adjacent water resources, also potentially affecting human health.

Effects of rare earth elements (REE) on terrestrial organisms: current status and future directions.

Rare Earth Elements (REE) are becoming increasingly important economically and highly exploited, thus contributing to REE increases in ecosystems. The ecotoxicological effects of REE on the terrestrial environment are, however, not fully understood and information on the biological effects of REE is urgently required for environmental risk assessments. In this review, studies and gaps in the existing scientific literature regarding the toxicological effects of REE on terrestrial organisms are presented. A total of 41 articles from the Web of Science database are discussed. La and Ce are the most studied elements, while little information is found concerning heavy REE. Most studies have been performed on plant species and few investigations are available for animals. Plant effects such as reduced mitotic index, germination and photosynthesis and antioxidant system enzyme alterations have been reported. Invertebrate effects include mortality, reproduction alterations and reduced locomotion. Based on the limited number of articles on terrestrial environment REE effects, this review highlights the need for more detailed studies in order to elucidate the effects associated with the REE hormesis and perform complete risk assessments with the establishment of safe REE usage limits.

Oxidative stress and metal homeostasis alterations in Danio rerio (zebrafish) under single and combined carbamazepine, acetamiprid and cadmium exposures.

Contaminants of emerging concern (CEC) are routinely detected in aquatic environments, especially pharmaceuticals, such as carbamazepine (CBZ), and neonicotinoid pesticides, like acetamiprid (ACT). CECs can interact with each other and with other legislated contaminants like Cd, resulting in unknown effects. Most studies evaluate only the effects of single contaminant exposures on aquatic biota. Therefore, the aim of the present study was to assess the effects of both single and combined CBZ, ACT and Cd exposures on zebrafish brain and liver oxidative stress parameters and metal homeostasis. The biomarkers catalase (CAT), glutathione-S-transferase (GST), total thiols (TOT), metallothionein (MT) and malondialdehyde (MDA) and the essential elements Ca, Cu, K, Na, Mg, Mn and Zn were evaluated after 96-hour static exposures. CBZ, ACT and Cd single (brain and liver) and combined (liver) treatments resulted in oxidative effects in both fish organs, also leading to metal (Ca, Mg, K, Mn, Zn and Cu) homeostasis alterations. ACT exposure resulted in the greatest adverse effects in the brain, while CBZ was the cause of major element homeostasis and oxidative stress alterations in the liver. Lower LPO levels were observed in the combined treatments compared to single treatments, suggesting interactions and contaminant effect attenuation. This study is the first to evaluate the initial effects of combined CBZ, ACT and Cd exposures in zebrafish, paving the way for further investigations concerning other biomarkers during longer exposure times.

Antioxidant system alterations and biological health status of earthworms following long-term exposure to antibiotic-contaminated poultry litter.

Poultry litter is widely applied as a fertilizer even though it is one of the main antibiotic sources to agricultural soils. Long-term sublethal effects (56 days) on the antioxidant system of Eisenia andrei earthworms following exposure to fluoroquinolone-contaminated poultry litter (enrofloxacin + ciprofloxacin) at 5.0, 10, and 20 g kg-1 were evaluated. The following biomarkers were assessed: superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), reduced glutathione (GSH), and a lipid peroxidation (LPO) proxy. Significant CAT and SOD increases, and a moderate positive correlation (ρ = 0.67, p < 0.05) between these enzymes was observed. Glutathione-S-transferase levels increased significantly at 10 g kg-1, while GSH exhibited a dose-dependent response at 5.0 mg kg-1 (4-106%), 10 mg kg-1 (28-330 %), and 20 mg kg-1 (45-472%). LPO levels exhibited a decreasing trend with increasing poultry litter concentrations of 8-170% (5.0 g kg-1), 7-104% (10 mg kg-1), and 3-6% (20 mg kg-1). A principal component analysis (PCA) highlighted increased SOD and CAT activities, possibly due to increased reactive oxygen species (ROS) concentrations. Biological health status assessments based on the biomarker response index indicate major alterations in the first month of exposure and becoming moderate in the second month. These findings indicate an antioxidant system attenuation trend. It is possible, however, that successive poultry litter applications may reduce the long-term recovery capacity of the evaluated biomarkers.

Long-term landfill leachate exposure modulates antioxidant responses and causes cyto-genotoxic effects in Eisenia andrei earthworms.

It is estimated that approximately 0.4% of the total leachate produced in a landfill is destined for treatment plants, while the rest can reach the soil and groundwater. In this context, this study aimed to perform leachate toxicity evaluations through immune system cytotoxic assessments, genotoxic (comet assay) appraisals and antioxidant system (superoxide dismutase - SOD; catalase - CAT, glutathione-S-transferase - GST; reduced glutathione - GSH and metallothionein - MT) evaluations in Eisenia andrei earthworms exposed to a Brazilian leachate for 77 days. The leachate sample contained high organic matter (COD - 10,630 mg L-1) and ammoniacal nitrogen (2398 mg L-1), as well as several metals, including Ca, Cr, Fe, Mg, Ni and Zn. Leachate exposure resulted in SOD activity alterations and increased CAT activity and MT levels. Decreased GST activity and GSH levels were also observed. Antioxidant system alterations due to leachate exposure led to increased malondialdehyde levels as a result of lipid peroxidation after the 77 day-exposure. An inflammatory process was also observed in exposed earthworms, evidenced by increased amoebocyte density, and DNA damage was also noted. This study demonstrates for the first time that sublethal effect assessments in leachate-exposed earthworms comprise an important tool for solid waste management.

Ecotoxicological evaluation of imazalil transformation products on Eisenia Andrei.

Data concerning the toxicity of the transformation products of some pesticides considered emerging contaminants are still incipient. This study aimed to evaluate acute (filter paper contact and avoidance test) and chronic (assays carried out in Red yellow Ultisoil) effects of the transformation products of the fungicide imazalil (IMZ) by heterogeneous photocatalysis (TiO2/UV) in Eisenia andrei. Some endpoints, i.e. biomass variation, disorder in reproduction and avoidance behavior, cytotoxicity (cell density and feasibility), and coelomic fluid (eleocytes and amoebocytes, immune cell) and antioxidant system (catalase (CAT) and glutathione S-transferase (GST)) changes were assessed. The studied degradation times (6, 18, 35 and 90 min) represented 70; 35; 10 and 0% of the initial IMZ concentration (5 mg L-1). No lethality and no significant difference in biomass variations were noted in the acute contact test. Decreased GST and increased CAT activities, as well as cell typing alterations and decreased cell viability and density detected at the 90-min degradation timepoint (PDT-90) indicate direct cytotoxic IMZ transformation product effects. In the avoidance test, no significant earthworm repulsion or attraction was observed. PDT-90 transformation products were responsible for losses in biomass and a reduction in the number of earthworm cocoons and juveniles present in soil. No cytotoxic effects were observed in the long term, although increased CAT and decreased GST activities were observed. Investigations on the relative toxicity of IMZ transformation products are relevant for future discussions regarding the environmental treatment, control and destination of this compound and its derivatives.

Lethal and long-term effects of landfill leachate on Eisenia andrei earthworms: Behavior, reproduction and risk assessment.

Leachate is difficult to biodegrade, and presents variable physical, chemical and biological characteristics, as well as high toxicological potential for soil, groundwater and water bodies. In this context, untreated leachate toxicity was evaluated through acute and chronic exposures in Eisenia andrei earthworms. Physico-chemical leachate characterizations indicate a complex composition, with high organic matter (COD - 10,634 mg L-1) and ammoniacal nitrogen (2388 mg L-1) concentrations. Metals with carcinogenic potential, such as Cr, As and Pb, were present at 0.60, 0.14 and 0.01 µg L-1, respectively and endocrine disrupting compounds were detected in estradiol equivalents of 660 ± 50 ng L-1. Acute tests with Eisenia andrei indicated an LC50 (72 h) of 1.3 ± 0.1 µL cm-2 in a filter paper contact test and 53.9 ± 1.3 mL kg-1 in natural soil (14 days). The EC50 in a behavioral test was estimated as 31.6 ± 6.8 mL kg-1, indicating an escape effect for concentrations ranging from 35.0 to 70.0 mL kg-1 and habitat loss from 87.5 mL kg-1 of leachate exposure. Chronic exposure (56 days) led to reproduction effects, resulting in a 4-fold decreased cocoon production and 7-fold juvenile decrease. This effect was mainly attributed to the possible presence of endocrine disrupting compounds. An estimated NOAEL of 1.7 mL L-1 and LOAEL of 3.5 mL L-1 were estimated for earthworms exposed to the assessed effluent. Extremely high-risk quotients (RQ ≥ 1) were estimated based on leachate application in irrigation. Thus, adequate municipal solid waste management is paramount, especially with regard to generated by-products, which can result in high toxicological risks for terrestrial organisms.

Fluoroquinolone-contaminated poultry litter strongly affects earthworms as verified through lethal and sub-lethal evaluations.

Poultry litter is one of the main sources of fluoroquinolones (FQs) in agricultural soils. In this study, our main goal was to investigate FQ-contaminated poultry litter effects on Eisenia andrei earthworms. To achieve this, acute and chronic tests covered several endpoints, such as avoidance, biomass, lethality, reproduction and changes to immune cells. FQs (enrofloxacin and ciprofloxacin) were determined in a poultry litter sample through high performance liquid chromatography with a fluorescence detector. The avoidance test indicates that poultry litter strongly repels earthworms, even at the lowest concentration (50 g kg-1). In the acute test, the lethal concentration of poultry litter to 50% of the earthworms (LC50), was estimated at 28.5 g kg-1 and a significant biomass loss (p < 0.05) occurred at 40 g kg-1. In the chronic test, a significant reproduction effect was observed at 20 g kg-1. Cell typing, density and feasibility indicated significant effects ranging from 5 to 20 g kg-1. A high risk quotient was estimated based on recommended poultry litter applications in field studies. Although FQ contamination in poultry litter and soils has been widely reported in previous studies, this is, to the best of our knowledge, the first toxicological assessment concerning earthworms exposed to FQ-contaminated poultry litter.

Contamination and oxidative stress biomarkers in estuarine fish following a mine tailing disaster.

BACKGROUND: The Rio Doce estuary, in Brazil, was impacted by the deposition of iron mine tailings, caused by the collapse of a dam in 2015. Based on published baseline datasets, the estuary has been experiencing chronic trace metal contamination effects since 2017, with potential bioaccumulation in fishes and human health risks. As metal and metalloid concentrations in aquatic ecosystems pose severe threats to the aquatic biota, we hypothesized that the trace metals in estuarine sediments nearly two years after the disaster would lead to bioaccumulation in demersal fishes and result in the biosynthesis of metal-responsive proteins. METHODS: We measured As, Cd, Cr, Cu, Fe, Mn, Pb, Se and Zn concentrations in sediment samples in August 2017 and compared to published baseline levels. Also, trace metals (As, Cd, Cr, Cu, Fe, Hg, Mn, Pb, Se and Zn) and protein (metallothionein and reduced glutathione) concentrations were quantified in the liver and muscle tissues of five fish species (Cathorops spixii, Genidens genidens, Eugerres brasilianus, Diapterus rhombeus and Mugil sp.) from the estuary, commonly used as food sources by local populations. RESULTS: Our results revealed high trace metal concentrations in estuarine sediments, when compared to published baseline values for the same estuary. The demersal fish species C. spixii and G. genidens had the highest concentrations of As, Cr, Mn, Hg, and Se in both, hepatic and muscle, tissues. Trace metal bioaccumulation in fish was correlated with the biosynthesis of metallothionein and reduced glutathione in both, liver and muscle, tissues, suggesting active physiological responses to contamination sources. The trace metal concentrations determined in fish tissues were also present in the estuarine sediments at the time of this study. Some elements had concentrations above the maximum permissible limits for human consumption in fish muscles (e.g., As, Cr, Mn, Se and Zn), suggesting potential human health risks that require further studies. Our study supports the high biogeochemical mobility of toxic elements between sediments and the bottom-dwelling biota in estuarine ecosystems.

Triclocarban affects earthworms during long-term exposure: Behavior, cytotoxicity, oxidative stress and genotoxicity assessments.

Triclocarban (TCC) is a contaminant of emerging concern widely applied as an antimicrobial in personal care products and introduced into the terrestrial environment through the application of biosolids (i.e., treated sewage sludge) in agriculture. Displaying the potential to bioaccumulate in the food chain and a high half-life in the soil, the presence of this compound in the environment may lead to potential ecological risks. In this context, TCC toxicity assessments in Eisenia andrei earthworms were carried out through acute, avoidance and chronic tests following cytotoxicity, antioxidant system, i.e. acatalase (CAT), glutathione-S-transferase (GST), glutathione (GSH), lipid peroxidation (LPO), and DNA damage (comet assay) evaluations. An LC50 of 3.3 ± 1.6 mg cm-2 in the acute contact test and an EC50 of 1.92 ± 0.31 mg kg-1 in the avoidance test during 72 h and 48 h, respectively, were obtained. The behavioral test indicates earthworm avoidance from 15.0 mg kg-1 of TCC. During chronic soil exposure, a 44% reduction in earthworm cell viability was observed after 14 days of exposure to 10 mg kg-1 TCC, while an increase in the percentage of amoebocyte cells also ocurred. Chronic exposure to TCC led to reduced CAT and GST activities, decreased GSH levels and increased LPO in exposed organisms. DNA damage was observed after 45 days from a 1 mg kg-1 dose of TCC. Therefore, TCC exhibits toxicological potential to Eisenia andrei earthworms, mainly during long-term exposures. This study provides mechanistic earthworm information towards understanding the environmental and human health implications of TCC exposure and draws attention to correct biosolid management.

Sub-lethal effects of the pesticide imazalil on the earthworm Eisenia andrei: reproduction, cytotoxicity, and oxidative stress.

Although considered an emerging contaminant and detected in the environment, the systematic and penetration fungicide imazalil ((RS)-1-(ß-allyloxy-2,4-dichlorophenylethyl) imidazole) has received relatively little scientific attention with regard to its possible negative effects in the environment. Only a few toxicological studies have assessed the potential environmental effect of imazalil and its impact on organisms. In this context, the aim of the present study is to evaluate the effects of different concentrations of the pesticide imazalil on the earthworm Eisenia andrei in acute contact and chronic tests in natural soil. Moreover, several endpoints, such as biomass loss or gain, reproduction, behavior, effects on immune system cells, and oxidative stress were also evaluated. Imazalil toxicity to E. andrei was determined by three approaches: a filter paper contact test (0, 0.16, 1.66, 16.6, 166 µg.cm-2), an avoidance (0, 0.1, 1, and 10 mg.kg-1), and a chronic test for 45 days (0, 0.01, 0.1, 1, and 10 mg.kg-1). All organisms exposed to the filter paper contact and chronic tests were submitted to two endpoint analyses: first, coelomic fluid collection by the extrusion method to determine density, viability, and cell type; second, oxidative stress assessments by determining GST and CAT enzymatic activities. This study allows for the conclusion that imazalil does not cause immediate earthworm death after exposure (LC50 > 166 µg.cm-1). However, due to several complementary factors, this compound may compromise earthworm health and lead to death, as E. andrei individuals did not avoid the contaminated soil, thus contributing to longer exposure periods and consequent cumulative damage to their systems. Decreased immunocompetent cellular viability (p < 0.05) and density (p < 0.05) in the chronic test are noteworthy, leading to susceptibility to exogenous factors, as well as irreversible cellular damage provoked by oxidative stress, such as cellular membrane rupture.

Lethal and sublethal effects of acetamiprid on Eisenia andrei: Behavior, reproduction, cytotoxicity and oxidative stress.

The neonicotinoid acetamiprid has been suggested as a worldwide substitute for organophosphates, due to its lower toxicity. The present study assessed several acetamiprid effects on Eisenia andrei earthworms in acute contact (ranging from 1.6 × 10-5 to 0.16 µg cm-2 acetamiprid), behavioral (0.1, 0.5, 1 mg kg-1) and chronic (0.001, 0.01, 0.05 and 0.1 mg kg-1 acetamiprid) assays carried out in natural soil. Reproduction, cytotoxicity (coelomocyte density and viability), immune cell typing (eleocytes and amoebocytes) and antioxidant defense system (glutathione (GSH), catalase (CAT) and glutathione S-transferase (GST)) responses were determined. The LC50 in the acute contact test was calculated as 1.86 × 10-2 µg cm-2. Acetamiprid concentrations of 0.5 and 1 mg kg-1 led to earthworm avoidance responses (NR = 61.09 ±â€¯10.01%) and habitat loss (NR = 78.02 ±â€¯12.03%), respectively. Reproduction was also affected, with a decreased number of cocoons and hatchlings per cocoon observed at 0.05 and 0.1 mg kg-1. Amoebocytes were the predominant immune system cells during the 15th and 30th assay days, while eleocytes were the main cells observed at the 45th day. CAT activities on the 30th and 45th day of exposure were increased at the lowest acetamiprid concentrations (0.001 and 0.01 mg kg-1) and decreased with increasing pesticide concentration (0.05 and 0.1 mg kg-1). Maximum GST activities and GSH levels were noted at 0.01 mg kg-1 acetamiprid. However, increasing concentrations led to GST inhibition, while GSH levels were maintained. A long-term acetamiprid exposure affected earthworm reproduction, behavior and immune and antioxidant systems, which could affect the ecological soil balance and, consequently, the entire food chain.

Spent lubricant oil-contaminated soil toxicity to Eisenia andrei before and after bioremediation.

Bioremediation is very efficient in biodegrading petroleum hydrocarbons. However, the decrease in these target contaminants in soils is not necessarily followed by a decrease in toxicity. The remaining contaminants can be enough to retain toxicity, while incomplete degradation of several compounds can generate sub-products, which can be even more toxic. In this context, the aim of this study was to assess acute and chronic toxicity in Eisenia andrei exposed to soil contaminated with 5% spent lubricant oil before and after 22 months of bioremediation in 150 L aerobic reactors. Applied bioremediation strategies were biostimulation (BIOS), bioaugmentation by adding mature compost from municipal solid waste (BIOA1) and bioaugmentation by adding non-mature compost from municipal solid waste (BIOA2). After 22 months, total petroleum hydrocarbons (TPH) were reduced 71% in BIOS and 73% in both BIOA1 and BIOA2. Polycyclic aromatic hydrocarbons (PAH) were reduced in about 98% in all treatments (BIOS, BIOA1 and BIOA2). At the 14th day of exposure, mortality rates were 7 ± 2, 20 ± 0, 75 ± 25, 93 ± 12 and 100 ± 0% for Eisenia andrei exposed to CONT (soil with no oil addition), BIOS, OLU (soil newly contaminated with 5% spent oil), BIOA1 and BIOA2, respectively. After 14 days, surviving specimens in both BIOS and OLU soils exhibited anatomic deformations, less biomass than the controls, and decrease in juvenile forms and coelomocytes. After 28 days, the mortality rate for BIOS and OLU soils increased to 97 and 100%, respectively. Therefore, even with a reduction of 71-73% for TPH and 98% for PAH, toxic effects remained in all soils bioremediated, probably due to the remaining hydrocarbons and/or hydrocarbon biodegradation products. The results indicate that both chemical analyses and toxicological monitoring are required to follow-up soil remediation progress.

Sublethal psychotropic pharmaceutical effects on the model organism Danio rerio: Oxidative stress and metal dishomeostasis.

Contaminants of emerging concern have become an important environmental problem, especially pharmaceutically active compounds (PhACs), since, after use, these drugs return to the environment, contaminating aquatic ecosystems. Some may display the ability to bioaccumulate and biomagnify throughout the food chain, leading to potential environmental and human deleterious effects which are, however, still largely unknown. In this context, the aim of the present study was to evaluate the effect of two psychotropic drugs commonly found in the environment, carbamazepine (CBZ) and clonazepam (CZP), both isolated and co-administrated, on oxidative stress biomarkers and essential metal homeostasis in Danio rerio fish specimens. No studies are available to data in this regard concerning CZP effects on fish. Reduced Glutathione (GSH), Metallothionein (MT), Catalase (CAT) and Glutathione S-Transferase (GST) were determined, as well as essential metals in fish liver, kidney and brains. Significant oxidative stress effects were observed for several biomarkers, where brain GST activity was the most affected, mainly with regard to CBZ exposure, while GST and CAT activity in the liver were downregulated in the co-administration mixture. In addition, dishomeostasis of several essential elements was detected in all analyzed organs, with a synergistic action of CBZ and CZP in brain, decreasing basal Mg, Al, K, Fe, Co, Ni and Cu levels in this organ, the target site for these drugs in humans. The brain organ was the most affected as observed by altered GST activity and metal dyshomeostasis concerning exposure to both compounds. These compounds, thus, present health risks to the aquatic biota, due to the evidenced deleterious effects and their constant release into the environment due to widespread use. Steps are needed to implement adequate legislation for risk analysis and decision-making in order to mitigate the effects of these emerging contaminants on aquatic ecosystem health.

Solar CPC pilot plant photocatalytic degradation of bisphenol A in waters and wastewaters using suspended and supported-TiO2. Influence of photogenerated species.

Photocatalytic degradation of bisphenol A (BPA) in waters and wastewaters in the presence of titanium dioxide (TiO2) was performed under different conditions. Suspensions of the TiO2 were used to compare the degradation efficiency of BPA (20 mg L(-1)) in batch and compound parabolic collector (CPC) reactors. A TiO2 catalyst supported on glass spheres was prepared (sol-gel method) and used in a CPC solar pilot plant for the photodegradation of BPA (100 µg L(-1)). The influence of OH·, O2 (·-), and h (+) on the BPA degradation were evaluated. The radicals OH· and O2 (·-) were proved to be the main species involved on BPA photodegradation. Total organic carbon (TOC) and carboxylic acids were determined to evaluate the BPA mineralization during the photodegradation process. Some toxicological effects of BPA and its photoproducts on Eisenia andrei earthworms were evaluated. The results show that the optimal concentration of suspended TiO2 to degrade BPA in batch or CPC reactors was 0.1 g L(-1). According to biological tests, the BPA LC50 in 24 h for E. andrei was of 1.7 × 10(-2) mg cm(-2). The photocatalytic degradation of BPA mediated by TiO2 supported on glass spheres suffered strong influence of the water matrix. On real municipal wastewater treatment plant (MWWTP) secondary effluent, 30 % of BPA remains in solution; nevertheless, the method has the enormous advantage since it eliminates the need of catalyst removal step, reducing the cost of treatment.