The use of integrative tools and multiple models for aquatic environmental quality assessment: a case study of the Mirim Lagoon, Southern Brazil.

This study performed toxicity assays with microalgae, microcrustaceans, and fish as well as evaluated biochemical and behavioral biomarkers in fish and microcrustaceans to assess the quality of the surface water of Mirim Lagoon, which belongs to one of the largest hydrographic basins in the world, located in southern Brazil. Three distinct sampling periods were chosen (January, March, and June 2022) based on the rice plantation dynamics which is the main activity surrounding the lagoon. In January, the plantation is irrigated; in March, the water is drained into the Mirim Lagoon, and July is the off-season. Concerning toxicity tests, there was significant inhibition in microalgae growth when exposed to water collected in March, but no mortality was observed for Ceriodaphia dubia, Daphnia magna, and Danio rerio. Regarding biomarkers, behavioral variables contributed more to the higher values of the Integrated Biomarker Response (IBR) index for both D. magna and D. rerio, in March. The Redundancy Analysis (RDA) indicated a correlation between the biomarkers for both organisms and abiotic parameters, mainly nutrients (total phosphorus and total nitrogen), thermotolerant coliforms, total solids, and turbidity. Spatially, there was no difference during monitoring, but the most significant ecotoxicological effects were observed in March. Multivariate analysis and the IBR index proved to be useful tools for monitoring of water bodies such as Mirim Lagoon.

The microbial profile of rivers and lagoons three years after the impact of the world's largest mining disaster (Fundão dam, Brazil).

The collapse of the Fundão tailings dam (Minas Gerais, Brazil) was the largest environmental disaster in Brazil's history and in the world mining industry. This disaster carried approximately 55 million m3 of iron ore tailings along the rivers and the lagoons of the Doce river basin. Although multiple studies assessed the impact on microbial communities in those rivers and lagoons right after the dam rupture, it is not known whether the microbiome in those environments remains impacted years after the disaster. Assessing the microbiome is very important to evaluate impacts and evaluate the health of the environment, due to the several ecological roles played by microorganisms. Here, we evaluated the impact of the dam failure on water and sediment bacteriome and archaeome by high-throughput next-generation sequencing. Samples were taken from two rivers and six lagoons during the dry and rainy seasons approximately three years post disturbance. The results showed a large number and abundance of microbial groups associated with the presence of heavy metals and mine tailings sediments. Some of these microorganisms were also reported in large abundance in the impacted rivers shortly after the Fundão dam rupture. Among the most abundant microorganisms in the Doce River, we can highlight the bacteria hgcI clade and the archaea Nitrososphera sp. in the water, and the bacteria Anaerolineaceae sp. in the sediment. These results suggest that the microbiome of the rivers and the lagoons in the Doce river basin remains severely impacted by the Fundão tailings dam failure even three years after the disaster. The presence of those microorganisms can also help to assess the occurrence of the Fundão dam sediment in other environments.

Toxicity of methylparaben to green microalgae species and derivation of a predicted no effect concentration (PNEC) in freshwater ecosystems.

Methylparaben (MeP) is one of the most used preservatives in the industry; however, the toxic effects on aquatic ecosystems are still poorly understood. Therefore, this study was conducted (1) to identify and compare the toxic effects of MeP on physiological parameters of different green microalgae species, using suitable mathematical models; and (2) to estimate a PNEC value for MeP in freshwater ecosystems, adopting either the deterministic or the probabilistic approaches. Toxicity tests were carried out with three green microalgae (Pseudopediastrum boryanum, Desmodesmus communis, Raphidocelis subcapitata), in which different endpoints such as growth rate, chlorophyll-a, and cell viability were measured and compared through the effective concentration which caused a response in x% of test organisms (ECx). ECx were obtained by adjusting different non-linear regression models for each microalgae dataset. Chlorophyll-a endpoint resulted in the lowest EC50 values, respectively 125, 81.2, 18.3 mg L-1 for D. communis, P. boryanum and R. subcapitata, showing R. subicapitata as the most sensitive, and D. communis as the most tolerant species to MeP (P < 0.05). PNEC was estimated from the present study and previous reports resulting in 5.7 and 65 µg L-1, respectively for the deterministic (PNECd) and the probabilistic (PNECp) approach. The development of chronic assays using test organisms from different ecological groups is encouraged to provide robust PNECp. In this meantime, we recommend the use of the estimated PNECd to support MeP risk assessments and policy formulation.

Impacts of the biocide chlorothalonil on biomarkers of oxidative stress, genotoxicity, and sperm quality in guppy Poecilia vivipara.

Chlorothalonil is a fungicide present in antifouling paints and other formulations used in agriculture, although studies have shown this chemical to be toxic to fish species. To clarify the deleterious effects of chlorothalonil for these non-target organisms, the present study evaluated the toxic effects of this biocide for the estuarine guppy Poecilia vivipara in terms of an acute mortality test (96 h) and the analysis of biomarkers of oxidative stress, genotoxicity, and sperm quality. The LC50 calculated for P. vivipara was 40.8 µg/L of chlorothalonil. For the analysis of biomarkers, fish were exposed (96 h) to 1 and 10 µg/L of chlorothalonil. It was observed that chlorothalonil alters the levels of pro- and antioxidants towards oxidative stress. In the gills, a negative effect on total antioxidant capacity (ACAP) was detected, while there was a reduction in the activity of glutathione S-transferase (GST) in the liver. However, levels of glutathione (GSH) and the activity and glutamate-cysteine-ligase (GCL) increased in both tissues, as a possible detoxification response. Following chlorothalonil exposure, oxidative damage measured by lipoperoxidation (LPO) significantly increased at the cellular level only (red blood cells (RBCs) and sperm cells). An increase in fluidity of membranes, reactive oxygen species concentration and micronuclei (MNs) incidence were also seen in RBCs. In sperm cells, LPO increased, while membrane and mitochondrial functionality as well as sperm motility decreased. Based on these results, chlorothalonil can be considered as a toxic compound for fish, causing genotoxicity and affecting the RBCs physiology and the fertility of males of P. vivipara.

Accumulation and effects of copper on aquatic macrophytes Potamogeton pectinatus L.: Potential application to environmental monitoring and phytoremediation.

This study investigated the ability of Potamogeton pectinatus L. to accumulate copper and its effects on plants. In accumulation tests, macrophytes were exposed (96 h) to different copper concentrations (0-1000 µM) and the metal was measured in media and plant tissues (roots, stems and leaves) to determine the bioconcentration factor (BCF). Plants accumulated high concentrations of copper in a dose-dependent manner and roots was the main organ for copper accumulation. However, the more copper increased in water, the more BCF values decreased. It may be due to either saturation of copper uptake or down-regulation of metal uptake by plants. In the physiological and morphological analyses, plants were kept (96 h) in Hoagland nutrient solution without copper, in full Hoagland solution (0.5 µM Cu) and in Hoagland medium with copper from 1 to 100 µM. The absence and the presence of copper above to 1 µM inhibited photosynthesis. Chlorophylls and carotenoid levels also decreased with the excess of copper, a fact that may have affected the photosystem II-dependent of chlorophyll and caused photosynthesis suppression. Only macrophytes at 10 µM Cu showed decrease in length and number of leaves on the 10th day of the test, when they died. Chlorosis and necrosis were observed in control groups and groups with extra copper, but not in Hoalgand group. Overall, the macrophyte P. pectinatus can be considered a suitable plant for monitoring environments contaminated by copper, based on results of copper accumulation in the plant, decrease in pigment concentration and presence of chlorosis and necrosis. However, values of BCF based on fresh water tissues was not proper to indicate the use of P. pectinatus for cleaning environments contaminated by copper.

Hematological and genotoxic biomarkers in a natural population of freshwater turtles Trachemys dorbigni (Duméril & Bibron, 1835) (Testudines: Emydidae) living in an urban area in Southern Brazil.

The present study aimed to report the morphometric and hematological indices and genotoxicity of a free-life population of D'Orbigny's slider turtles (Trachemys dorbigni) living in an urban area in Southern Brazil. For that, 16 specimens were randomly captured in an urban canal that receives irregular releases of wastewater. Biometrics and external visual changes were analyzed, such as turtle shell deformities, and the presence of parasites. Blood samples were collected to evaluate the hematological profile and the presence of micronuclei and other erythrocyte nuclear abnormalities as potential mutagenic and genotoxic effects. Water physicochemical parameters were also measured. Organisms with ectoparasites (31.25%) and small carapace deformations (56.25%) were observed, but maximum carapace length and weight were considered normal for the species according to the literature. The blood profile indicated low hemoglobin and hematocrit and a high number of total leukocytes, particularly eosinophils which characterize parasitic infections. A frequency of 0.12% for the micronucleus was considered basal, but the frequency of other erythrocyte abnormalities was evident, mainly of blebbed nuclei (63.79%), indicating chromosomal damage in the early stage. The results of this study suggest that natural populations of chelonian inhabiting urbanized areas are impacted by anthropogenic activities in the surrounding environment. Furthermore, it provides comprehensive data which can serve as a comparative model for environmental monitoring studies involving turtles.

Glyphosate and its formulated product Roundup Transorb R® affect locomotor activity and reproductive and developmental parameters in Jenynsia lineata fish: An intergenerational study.

Glyphosate is the most widely utilized herbicide worldwide due to its effectiveness in controlling agricultural weeds. However, its persistence in aquatic ecosystems has raised concerns about the well-being of non-target organisms such as fish. This study aimed to evaluate the effects of chronic exposure (21 days) to glyphosate or its formulated product Roundup Transorb R®, at an environmentally relevant concentration permitted by regulations in certain countries (65 µg/L of glyphosate), on the locomotor activity and reproductive success of the fish Jenynsia lineata, as well as on the morphology/development and locomotor activity of its offspring, as intergenerational effects. Neither the pure nor formulated herbicide altered the distance traveled and velocity of adult fish exposed to the herbicide (F0), but they negatively affected reproductive success, decreasing the percentage of positive response to the presence of the female, reducing the number of gravid females, causing abortions, and lowering offspring survival (F1). In the F1 generation, a decrease in weight and length was noted along with developmental abnormalities in both treatment groups (pure or formulated glyphosate), with the formulation causing more harm. Observed developmental abnormalities included muscle atrophy, ascites, pigmentary disorders, vertebral agenesis, spinal deviation, and exophthalmia. Furthermore, parental exposure to pure glyphosate led to an increase in the distance traveled and velocity of F1 (hyperlocomotion), whereas exposure to the formulated product resulted in a decrease in these behaviors (hypolocomotion) of F1. These findings highlight the toxic effects of glyphosate at very low concentrations, although varying between pure and formulated, and demonstrate the intergenerational consequences of herbicide exposure, underscoring the risk to the survival of fish offspring in glyphosate-contaminated environments.

Effects of the Pesticide Carbofuran on Two Species of Chlorophyceae (Desmodesmus communis and Pseudopediastrum boryanum) and Their Pesticide Bioremediation Ability.

Carbofuran is one of the most toxic broad-spectrum pesticides. We evaluated the effects of carbofuran on two species of microalgae, Pseudopediastrum boryanum and Desmodesmus communis, through measurements of cell viability, biomass, chlorophyll content, and the production of reactive oxygen species (ROS). The ability of these algae to remove carbofuran dissolved in the media was also determined. For the evaluations, both microalgae species were exposed to carbofuran (FURADAN 350 SC®) at concentrations of 100, 1000, and 10,000 µg L-1 for 7 days. Algae cell viability and chlorophyll-a concentration were not affected by the presence of carbofuran. Both species grew when exposed to the pesticide; however, the microalgae D. communis grew less than its respective control when exposed to the highest concentration (10,000 µg L-1 of carbofuran), indicating an adverse effect of the pesticide on this species. A significant increase in ROS production was observed in D. communis and P. boryanum when exposed to the highest concentration tested. The microalgae P. boryanum completely removed carbofuran in the media within 2 days, regardless of the concentration, whereas D. communis achieved the same result only after 5 days of exposure. Growth inhibition was observed only until the disappearance of carbofuran from the media. The present study suggests the use of microalgae, mainly P. boryanum, as potential tools for the remediation of environments contaminated by carbofuran because of their resistance to the insecticide and their ability to remove it rapidly from water. Environ Toxicol Chem 2024;43:926-937. © 2023 SETAC.

Effects of the herbicide glyphosate on fish from embryos to adults: a review addressing behavior patterns and mechanisms behind them.

The use of agrochemicals has grown in recent years following the increase in agricultural productivity, to eliminate weeds that can compromise crop yields. The intensive use of these products combined with the lack of treatment of agricultural wastewater is causing contamination of the natural environments, especially the aquatics. Glyphosate [N-(phosphonomethyl) glycine] is the most commonly used herbicide in agriculture worldwide. Studies have shown that this compound is toxic to a variety of fish species at the concentrations of environmental relevance. Glyphosate-based herbicides can affect fish biochemical, physiological, endocrine, and behavioral pathways. Changes in behaviors such as foraging, escaping from predators, and courtship can compromise the survival of species and even communities. The behavior patterns of fish has been shown to be a sensitive tool for risk assessment. In this sense, this review summarizes and discusses the toxic effects of glyphosate and its formulations on the behavior of fish in different life stages. Additionally, behavioral impairments were associated with other negative effects of glyphosate such as energy imbalance, stress responses, AChE inhibition, and physiological and endocrine disturbances, which are evidenced and described in the literature. Graphical abstract.

Differential Effects of Atrazine on Chlorophyceae Species and Association with Morphology, Photosynthesis, Chlorophyll Content, and Glutathione-S-Transferase Activity.

Atrazine is a herbicide widely used in the control of weeds in crops such as corn, sugar cane, and sorghum. It is often found in aquatic environments, where it can potentially endanger nontarget organisms such as microalgae. The present study evaluated atrazine toxicity to seven different species of Chlorophyceae and the tolerance of the species to the herbicide was related to morphological, photosynthetic, chlorophyll-a content and the activity of the glutathione-S-transferase enzyme (GST). The comparison of median effect concentration (EC50) values for growth inhibition indicates higher toxicity of atrazine for Pseudopediastrum boryanum and Desmodesmus communis, intermediate toxicity for Ankistrodesmus densus, Chlamydomonas puliminiorfes, and Raphidocelis subcapitata, and lower toxicity for Kirchneriella lunaris and Ankistrodesmus falcatus (EC50: 38, 42, 66, 103, 248, 1004, and 1585 µg L-1 atrazine, respectively). Principal component analysis (PCA) with algal characteristics suggested that the atrazine-sensitive algae P. boryanum and D. communis were positively associated with photosynthetic levels and negatively associated with GST activity and chlorophyll-a concentration. The PCA also suggested that the atrazine-tolerant algae A. falcatus and K. lunaris were positively associated with morphological parameters, where the larger the cell size, the more tolerant. Although it is difficult to associate a single characteristic of algae as the key factor determining the tolerance to atrazine, results presented in this work indicate that the cell area, the photosynthetic parameters (mainly saturating irradiance), chlorophyll-a content, and the biotransformation by GST in combination may be potential predictors for the differential tolerance of Chlorophyceae species to the herbicide. Environ Toxicol Chem 2022;41:1675-1685. © 2022 SETAC.

Impacts of tailings of Fundão dam (Brazil) rupture on marine fish: Metals bioaccumulation and physiological responses.

This study evaluated the impacts of the mining tailings after the rupture of the Fundão dam on fish communities on the Atlantic Ocean southeast coast. Four sample collections were carried out over two years (2018-2020), in seasonal periods. Omnivorous/herbivorous and carnivorous fish were collected for analysis of metal bioaccumulation, multibiomarkers of environmental contamination and histopathology. Metal bioaccumulation was stronger correlated in carnivorous fish in the dry-2018 collection, besides higher activity of antioxidant enzymes, energy metabolism and higher morphological damage; however, there was less oxidative damage and less metallothioneins induction, and these variables were strongly associated with the wet-2020 collection. In a temporal view, it was possible to observe a reduction in metal levels in fish, except in the mouth of the Doce River. These events can be explained by seasonal natural events, which tend the resuspension and boost metal levels, mainly in the mouth region during the rainy season.

Effects of anti-inflammatory diclofenac assessed by toxicity tests and biomarkers in adults and larvae of Danio rerio.

The entrance of the anti-inflammatory diclofenac in water bodies is a consequence of inappropriate use, incorrect disposal, and inefficiency of wastewater treatment plants (WWTPs) in removing this drug from sewage, among others. Effects of diclofenac on non-target aquatic organisms still need to be clarified. The objective of this work was to evaluate the toxic effects of the diclofenac on larvae and adults of Danio rerio. The LC50 values were 5.49 mg/L and 5.22 mg/L for the adult and larvae, respectively. A set of biochemical and genotoxic biomarkers were evaluated in fish exposed to an environmentally relevant concentration of 2 µg/L DCF and a no observed effect concentration (NOEC) of 3 mg/L diclofenac. At the NOEC, an increase in activities of glutathione-S-transferase (GST) enzyme and an increase in ATP binding cassette (ABC) transporters in gills of adult fish was observed; also, an increase in lipoperoxidation (LPO) was seen in the gills of adults and whole larvae. These results indicate that diclofenac activates the fish detoxification processes and may affect fish health.

A scientometric analysis of ecotoxicological studies with the herbicide atrazine and microalgae and cyanobacteria as test organisms.

Atrazine (ATZ) is one of the most widely used herbicides in the world. A scientometric study was conducted to analyze the evolution of research on ATZ. The study also looked at the use of microalgae and cyanobacteria as biological models for toxicity tests during the period from 1959 to 2019, in the category of toxicology of Web of Science. The results show an increase in the number of scientific publications mainly in the USA, Canada, and China. The majority of papers was published in journals with high impact factors, demonstrating the relevance of such studies. About 83% of the studies aimed to evaluate the effect of ATZ on non-target organisms. From those, 7.5% included microalgae and cyanobacteria. The majority of them worked with chlorophyceae to perform toxicity bioassays of ATZ and analyze its sublethal effects. The gaps identified by this analysis included a small number of collaborations between research groups from different countries; the number of studies with terrestrial organisms, which are larger in comparison to aquatic organisms; and the fact that none of the studies with ATZ and microalgae was performed in the field. These findings can point out to researchers and funding agencies the gaps in knowledge on the toxic effects of ATZ and guide the development of new research projects as well as environmental policies.

Copper exposure alters the metabolism of the blue crab Callinectes sapidus submitted to osmotic shock.

Copper (Cu) is an essential metal capable to alter many metabolic and physiological processes in animal species, depending on the environmental concentration and salinity. The present study evaluated the effects of Cu exposure on the metabolism of the blue crab Callinectes sapidus under different osmotic situations. Crabs were acclimated at two different salinities conditions (30 and 2). Subsequently, they were exposed to Cu during 96 h at each salinity and under hypo-osmotic shock. Results demonstrated that Cu exposure increased whole-body oxygen consumption. In addition, the activity of LDH decreased while citrate synthase increased in anterior gills from animals submitted to hypo-osmotic shock. This scenario indicates extra stress caused by sudden environmental osmotic changes, as commonly observed in estuarine environments, when combined with copper exposure. Therefore, the activity of LDH and citrate synthase enzymes might be sensitive indicators for aquatic toxicology studies approaching Cu contamination in estuarine environments.

Zebrafish (Danio rerio) ability to activate ABCC transporters after exposure to glyphosate and its formulation Roundup Transorb®.

The emergent demand for food production has increased the widespread use of pesticides, especially glyphosate-based herbicides as they can protect different types of crops, especially transgenic ones. Molecules of glyphosate have been found in water bodies around the world, and its presence can cause negative effects on non-target organisms, such as fish. Glyphosate toxicity appears to be systemic in fish but does not affect their organs equally. Also, its formulations can be more toxic than pure glyphosate. In this sense, we investigated if these variations in toxicity could be related to ATP binding cassette subfamily C (ABCC) transporters and the cellular detoxification capacity, following exposure to herbicides. Thus, adults of Danio rerio were exposed (24 and 96 h) to glyphosate and Roundup Transorb® (RT) at an environmental concentration of 0.1 mg/L, and the activity of ABCC proteins and gene expression of five isoforms of ABCC were analyzed. Glyphosate and RT exposure increased ABCC protein activity and gene expression up to 3-fold when compared to controls, indicating the activation of detoxification mechanisms. Only in the brain of D. rerio, the exposure to RT did not stimulate the activity of ABCC proteins, neither the expression of genes abcc1 and abcc4 that responded to the exposure to pure glyphosate. These results may suggest that the brain is more sensitive to RT than the other target-tissues since the mechanism of detoxification via ABCC transporters were not activated in this tissue as it was in the other.

Effects of Roundup formulations on biochemical biomarkers and male sperm quality of the livebearing Jenynsia multidentata.

Roundup® formulations are the most consumed glyphosate-based herbicides in the world. When applied, they may reach water bodies and exert toxicity toward non-target species. This study evaluated and compared the effects of different variations of Roundup on biochemical biomarkers as oxidative parameters and Acethylchorinesterase (AChE) activity, and sperm quality of the livebearing Jenynsia multidentata. Fish were acutely (96 h) exposed to Roundup Original® (RO), Roundup Transorb® (RT) and Roundup WG® (RWG) at 0.0, 0.5, 1 and 5 mg L-1 of nominal glyphosate. The highest mortality (60%) was observed for fish exposed to RT at the highest concentration tested and at 0.5 mg L-1 non-mortality was observed, so this concentration was chosen for the experiments. Fish exposed to RO and RT (24 and 96 h) presented a state of oxidative imbalance, which caused lipid peroxidation (LPO) in their livers. Oxidative stress was more severe in RO treatment, which may be resulted in the highest hepathosomatic index at 96 h. However, fish exposed to RT presented a marked inhibition of AChE activity from membrane cells of muscle and brain tissues. Sperm quality was investigated in livebearing exposed (24 and 96 h) to the three formulations. Spermatozoa motility and concentration were affected by all formulations. Overall, Roundup formulations are harmful to the fish J. multidentata at 0.5 mg L-1 of glyphosate; however, mechanisms and potential of toxicity are different between formulations. The J. multidentata also represents a sensitive species and a good regional bio-monitor.

Impaired regulation of divalent cations with acute copper exposure in the marine clam Mesodesma mactroides.

The mechanism of copper (Cu) toxicity in marine invertebrates remains unclear. Therefore, marine clams (Mesodesma mactroides) were exposed (96h) to a concentration of dissolved Cu (1.6µmolL(-1)) inducing 10% mortality in sea water (30ppt). After in vivo exposure, tissue Cu accumulation (hemolymph, gill and digestive gland); hemolymph ionic (Na(+), K(+), Mg(2+) and Ca(2+)) and osmotic concentrations; tissue (gill and digestive gland) ionic concentration, enzyme (Na(+),K(+)-ATPase and carbonic anhydrase) activity, and oxygen consumption; and whole-body oxygen consumption were analyzed. Succinate dehydrogenase activity was evaluated in mitochondria isolated from gills and digestive gland and exposed (1h) in vitro to different concentrations of dissolved Cu (0.8, 7.7 and 78.7µmolL(-1)). In vivo exposure induced Cu accumulation in hemolymph, gills and digestive gland; increased Mg(2+) and decreased Ca(2+) concentration in hemolymph; decreased Mg(2+) concentration, increased Na(+),K(+)-ATPase activity and reduced carbonic anhydrase activity in gills; decreased Mg(2+) concentration, increased Ca(2+) concentration and increased Na(+),K(+)-ATPase activity in digestive gland; and reduced gill, digestive gland and whole-body oxygen consumption. Succinate dehydrogenase activity was inhibited after in vitro exposure to 78.7µmolL(-1) Cu. These findings indicate that Cu is an ionoregulatory toxicant in the marine clam M. mactroides. However, toxicity is related to disturbances in regulation of divalent cations (Mg(2+) and Ca(2+)) without effect on regulation of major monovalent cations (Na(+) and K(+)), as opposed to that observed in osmoregulating invertebrates exposed to Cu. However, other mechanism(s) of toxicity cannot be ruled out. Future studies must be performed to evaluate the consequence of the Cu-induced respiratory disturbances observed in M. mactroides.

Effects of glyphosate on cholinesterase activity of the mussel Perna perna and the fish Danio rerio and Jenynsia multidentata: in vitro studies.

Although the herbicide glyphosate [N-(phosphonomethyl)glycine] is not classified as an acethylcholinesterase inhibitor, some studies have reported reduction in the acethylcolinesterase activity after in vivo exposure to both its pure form and its commercial formulations. Considering this controversy, the objective of the present study was to investigate, in vitro, the effects of glyphosate exposure on cholinesterase activity of the brown mussel Perna perna and of two fish species: zebrafish Danio rerio and onesided livebearer Jenynsia multidentata. For this purpose, samples of different tissues (brain and muscle for fish; gills and muscle for mussel) were homogenized and pre-incubated with different glyphosate concentrations before cholinesterase activity determination. Results demonstrated that cholinesterase from different fractions of all species tested was inhibited by glyphosate. The concentrations of glyphosate that inhibits 50% of cholinesterase activity (IC50) ranged from 0.62 mM for P. perna muscle to 8.43 mM for J. multidentata brain. According to this, cholinesterase from mussel seems to be more sensitive to glyphosate exposure than those from the fish D. rerio and J. multidentata.

Acute toxicity, accumulation and tissue distribution of copper in the blue crab Callinectes sapidus acclimated to different salinities: in vivo and in vitro studies.

In vivo and in vitro studies were performed to evaluate acute toxicity, organ-specific distribution, and tissue accumulation of copper in Callinectes sapidus acclimated to two different experimental salinities (2 and 30 ppt). Blue crabs were quite tolerant to copper. Acute dissolved copper toxicity (96-h LC(50) and its corresponding 95% confident interval) was higher at salinity 2 ppt (5.3 (3.50-8.05) µM Cu) than at 30 ppt (53.0 (27.39-102.52) µM Cu). The difference between salinities can be completely explained based on the water chemistry because it disappeared when 96-h LC(50) values were expressed as the free Cu(2+) ion (3.1 (1.93-4.95) µM free Cu at 2 ppt versus 5.6 (2.33-13.37) µM free Cu at 30 ppt) or the Cu(2+) activity (1.4 (0.88-2.26) µM Cu activity at 2 ppt versus 1.7 (0.71-4.07) µM Cu activity at 30 ppt). The relationships between gill Cu burden and % mortality were very similar at 2 and 30 ppt, in accord with the Biotic Ligand Model. In vivo experiments showed that copper concentration in the hemolymph is not dependent on metal concentration in the surrounding medium at either experimental salinity. They also showed that copper flux into the gills is higher than into other tissues analyzed, and that anterior and posterior gills are similarly important sites of copper accumulation at both experimental salinities. In vitro experiments with isolated-perfused gills showed that there is a positive relationship between copper accumulation in this tissue and the metal concentration in the incubation media for both anterior and posterior gills. A similar result was observed at both low and high salinities. Furthermore, in vitro experiments showed that copper accumulation in posterior gills is also positively and strongly dependent on the incubation time with copper. Gill copper accumulation occurred at a lower rate in the first 2h of metal exposure, increasing markedly after this "steady-state" period. This finding was corroborated by a significant increase in copper influx to the gill perfusate (corresponding to crab hemolymph) after this time, measured using (64)Cu. In vivo, after uptake from solution, (64)Cu was primarily accumulated in the gills and the rest of the body rather than in the hemolymph, hepatopancreas, or other internal tissues. Overall, the present findings indicate that gills are a key target organ for copper accumulation, as well as an important biological barrier against the excessive uptake of copper into the hemolymph and the subsequent distribution of this metal to internal organs of the blue crab.