Electrochemical impedance biosensor for detection of saxitoxin in aqueous solution.

Saxitoxin is a cyanotoxin which is very harmful to human health; the concentration limit in drinking water is only 3 µg/L. Therefore, a simple, fast, sensitive, low-cost, and specific method for its detection, quantification, and monitoring in water bodies is needed to avoid adverse effects on animal and human health. In this work, we developed an electrochemical impedimetric biosensor using a specific aptamer as recognition element for saxitoxin detection. This method allies the superior sensing characteristics of aptamers with the nondestructive, label-free, and easy working principles of the electrochemical impedance technique. The device presented sensitivity for detecting saxitoxin concentrations above 0.3 µg/L, with high selectivity in negative control experiments, demonstrating a promising alternative for water toxin detection.

Synthesis, characterization and toxicological evaluation of Cr2O3 nanoparticles using Daphnia magna and Aliivibrio fischeri.

Chromium III oxide (Cr2O3) nanoparticles (NPs) are used in pigments for ceramics, dyes, paints and cosmetics. However, few studies addressing the toxic potential of these NPs have been reported in the literature. Thus, this research aimed to evaluate the acute and chronic effects of Cr2O3 NPs through acute toxicity tests with Daphnia magna and Aliivibrio fischeri and chronic toxicity tests with Daphnia magna. Cr2O3 NPs were synthesized by the sol-gel method and characterized through TEM, X-Ray diffraction (XRD), zeta potential (ZP) and surface area analysis. In the acute toxicity tests the EC(50,48h) value obtained with D. magna was 6.79 mg L(-1) and for A. fischeri the EC(50,15min) value was 16.10 mg L(-1) and the EC(50,30min) value was 12.91 mg L(-1). Regarding the chronic toxicity tests with D. magna, effects on longevity (OEC=1.00 mg L(-1)), reproduction (OEC=1.00 mg L(-1)) and growth (OEC=0.50 mg L(-1)) were observed. On the SEM and TEM images, ultrastructural alterations in the organelles of exposed organisms were also observed. Thus, toxicological studies with NPs are of great importance in order to reduce the risk of environmental contamination.

Evaluation of Cytotoxicity and Cell Death Induced In Vitro by Saxitoxin in Mammalian Cells.

Since the cyanotoxin saxitoxin (STX) is a neurotoxin and induces ecological changes in aquatic environments, a potential risk to public and environmental health exists. However, data on STX-mediated cytotoxic and genotoxic effects are still scare. In order to gain a better understanding of the effects of this toxin, the cytotoxic and genotoxic potential of STX was examined in two mammalian cell lines. Neuro 2A (N2A), a neuroblastoma mouse cell line, and Vero cell line, derived from Vero green monkey kidney cells, were exposed to several concentrations of STX ranging from 0.5 to 64 nM to determine cell viability, induction of apoptosis (DNA fragmentation assay), and formation of micronuclei (MN) (cytokinesis-block micronucleus assay; CBMN) following 24 h of incubation. The half maximal effective concentration (EC50) values for STX calculated in cell viability tests were 1.01 nM for N2A and 0.82 nM for Vero cells. With increasing STX concentration there was evidence of DNA fragmentation indicating apoptosis induction in Vero cells with a 50% increase in DNA fragmentation compared to control at the highest STX concentration tested (3 nM). The results demonstrated no significant changes in the frequency of micronucleated binucleated cells in N2A and Vero cells exposed to STX, indicating the absence of genotoxicity under these test conditions. There was no apparent cellular necrosis as evidenced by a lack of formation of multinucleated cells. In conclusion, data reported herein demonstrate that STX produced death of both cell types tested through an apoptotic process.

Induction of micronucleus of Oreochromis niloticus exposed to waters from the Cubatão do Sul River, southern Brazil.

In an effort to characterize the pollution of surface waters by potentially genotoxic agents, this study aimed at assessing the frequency of micronucleated (MN) erythrocytes of the fish species, Oreochromis niloticus, from the Cubatão do Sul River. This river is the source of drinking water for the region of Florianópolis, capital of Santa Catarina State, Brazil. Negative control fish showed low frequency of MN, ranging between 0.49‰ and 0.90‰. Positive control (potassium dichromate 2.5 mg/L) organisms showed high MN frequency (16.82-17.25‰). The MN frequency increased along the river (Site 1--1.24‰ winter 2011; Site 4--9.76‰ summer 2011). Based on the observation of elevated MN erythrocytes frequency in O. niloticus exposed to water samples from along the river course, we conclude that the complex environmental mixtures of water from the Cubatão do Sul River have genotoxic potential. This genotoxicity most likely originated from agricultural runoff and domestic effluents released without treatment, based on the evidence from literature data and a survey in the region. This study provides a scientific basis for future studies regarding the genotoxicity of complex environmental mixtures in natural environments.

Can the sonication of polystyrene nanoparticles alter the acute toxicity and swimming behavior results for Daphnia magna?

The seemingly ubiquitous presence of plastic debris led to a greater focus on micro- and nanoplastics research derived from the degradation process of macroplastics. The ingestion and consequent accumulation of plastics on the biota are the main concerns. Researchers strive to make assay conditions as close as possible to those of the environment. In this regard, sonication can be applied to de-agglomerate the plastic particles, but this may alter significantly their toxicity. The aim of this study was to understand the effects of the sonication process on the acute toxicity and swimming behavior of polystyrene nanoparticles using Daphnia magna as the test organism. The results show a 2-fold reduction in the acute toxicity after the sonication process; the EC50 of the PSNP-NS was 1.28 ± 0.17 mmol while for PSNP-S the EC50 was 2.77 ± 0.32 mmol, possibly through the formation of an eco-corona on the nanoplastic surface, formed from the ions dispersed in the medium or proteins secreted by the test organisms. The mean swimming distance was reduced when compared to the control group for both the PSNP-S and PSNP-NS. This is the first research stating the toxicological differences between sonicated and non-sonicated polystyrene nanoparticle samples using Daphnia magna as test organism.

Toxicological effects of AgNPs on duckweed (Landoltia punctata).

Silver nanoparticles (AgNPs) are widely applied in several types of products since they act as a biocide. However, their high level of release into the environment can bring risks to ecosystems. Thus, the toxicity of AgNPs toward duckweed (Landoltia punctata) was investigated by monitoring the growth rate inhibition and the effect on the photosynthetic metabolism through morphological and ultrastructural analysis. The AgNPs were characterized by transmission electron microscopy and the effective diameter (dynamic light scattering) and zeta potential were determined. Plants were grown according to the environmental conditions recommended in ISO/DIS 20079 and then exposed to different concentrations of AgNPs. Inhibition of the growth rate was measured based on the EC50 and changes in the morphology, cellular structures and photosynthetic pigments were evaluated along with the silver accumulation. Although the results showed low growth inhibition when compared to other studies, significant damage to the ultrastructure, decreases in the photosynthetic pigments and starch grains, an increase in the phenolic compounds and physiological changes, such as a loss of color, were observed. Moreover, the accumulation of silver ions was noted and this could lead to bioamplification in consumer organisms, since duckweed belongs to the first level of the food chain.

Comparison of cytotoxicity of α-Al2O3 and η-Al2O3 nanoparticles toward neuronal and bronchial cells.

The role of the crystalline structure on the toxicity of two phases of Al2O3 NPs, alpha (α-Al2O3 NPs) and eta (η-Al2O3 NPs), was investigated in this study. Different techniques were employed for the characterization of the Al2O3 NPs and multiple toxicological endpoints were used to assess the toxicity toward mouse neuroblastoma (N2A) and human bronchial epithelial (BEAS-2B) cells. Based on the results of the multiple toxicological endpoints, revealed differences in the toxic potential results for α-Al2O3 NPs and η-Al2O3 NPs, with the latter showing a more pronounced effect. These effects could be due to the high uptake of the η-Al2O3 NPs in the cytoplasmic vesicles, as evidenced by TEM and ICP-MS. Hence, the results demonstrate the potential toxicity of both α-Al2O3 NPs and η-Al2O3 NPs, although the N2A and BEAS-2B cells showed greater susceptibility toward η-Al2O3 NPs. Thus, our study demonstrates the important role of the crystalline structure in relation to the nanotoxicity of Al2O3 NPs.

Correlation between acute toxicity for Daphnia magna, Aliivibrio fischeri and physicochemical variables of the leachate produced in landfill simulator reactors.

Due to the diversified nature of municipal solid waste and the different stages of its decomposition, the formed leachates result in a complex chemical mixture with toxic potential. These chemicals can cause environmental problems, such as the contamination of surface or groundwater, thus affecting the balance of aquatic ecosystems. The aim of our study was to evaluate the acute toxicity of leachates in Daphnia magna and Aliivibrio fischeri and to identify the main physicochemical variables that influence the toxicity of the landfill leachates produced in reactors within pilot simulations. Acute toxicity tests carried out on D. magna and A. fischeri showed that the leachates produced inside the reactors are highly toxic, presenting EC5048h < 1% for D. magna and EC5015min < 12% for A. fischeri. This result indicates that microcrustaceans are more sensitive to leachates, making them more suitable to our study. Pb showed the highest correlation with EC5048h, suggesting that Pb is the main chemical variable indicative of toxicity for the conditions of the experiment. In smaller scale, phosphate (PO43-) and nitrate (NO3-) were the macronutrients that most influenced the toxicity. Clearly, this correlation should be viewed with caution because the synergistic effects of this complex mixture are difficult to observe.

Comparative study of Domoic Acid and Okadaic Acid induced-chromosomal abnormalities in the Caco-2 cell line.

Okadaic Acid (OA) the major diarrheic shellfish poisoning (DSP) toxin is known as a tumor promoter and seems likely implicated in the genesis of digestive cancer. Little is known regarding genotoxicity and carcinogenicity of Domoic Acid (DA), the major Amnesic Shellfish Poisoning (ASP) toxin. Both OA and DA occur in seafood and are of human health concerns. Micronuclei (MN) arise from abnormalities in nuclear division during mitosis due to a failure of the mitotic spindle or by complex chromosomal configurations that pose problems during anaphase. In order to evaluate the ability of okadaic acid (OA) and domoic acid (DA) to induce DNA damage we performed the micronucleus assay using the Caco-2 cell line. To discriminate between a clastogenic or aneugenic effect of OA and DA, the micronucleus assay was conducted by cytokinesis-block micronucleus assay using cytochalasin B with Giemsa staining and/or acridine orange staining, in parallel to fluorescence in situ hybridization (FISH) using a concentrated human pan-centromeric chromosome paint probe. Our results showed that OA and DA significantly increased the frequency of MN in Caco-2 cells. The MN caused by OA are found in mononucleated cells and binucleated cells, whereas those caused by DA are mainly in binucleated cells. The results of FISH analysis showed that OA induced centromere-positive micronuclei and DA increased the percentage of MN without a centromeric signal. In conclusion, both OA and DA bear mutagenic potential as revealed in Caco-2 cells by induction of MN formation. Moreover, OA induced whole chromosome loss suggesting a specific aneugenic potential, whereas DA seems simply clastogenic. At present, one cannot rule out possible DNA damage of intestinal cells if concentrations studied are reached in vivo, since this may happen with concentrations of toxins just below regulatory limits in case of frequent consumption of contaminated shell fishes.

Synthesis, characterization and toxicological evaluation of a core-shell copper oxide/polyaniline nanocomposite.

The newest generation of copper oxide NPs (CuO NPs) is the CuO core-shell (CS), which has potential applications in several areas (e.g., electronics and paint) and is able to provide a greater service life due to its coating; however, its toxicity is not fully understood. The objective of this study was to synthesize, characterize and evaluate the aquatic toxicology of CuO NPs and CuO core-shells through acute and chronic toxicity tests with the freshwater microcrustaceans Daphnia magna and to evaluate its acute toxicity with the marine bioluminescent bacteria Vibrio fischeri. The NPs were synthesized by direct thermal decomposition after being coated as a CS with polyaniline (PANI). With respect to acute toxicity with D. magna, the CuO NPs and CS CuO/PANI presented EC50 values of 0.32 mg L(-1) and 0.48 mg L(-1), respectively. For the tests with V. fischeri, the CuO NPs (EC50-15 min=7.79 mg L(-1)) exhibited behavior similar to that of the CS CuO/PANI (EC50-15 min=9.05 mg L(-1)) after 15 min of exposure. Regarding chronic toxicity, both forms showed a statistically significant effect (p<0.05) on the growth and reproduction parameters. Based on the characterization and toxicity results, it can be concluded that both forms of CuO were toxic and presented similar behaviors during the acute tests; however, after 21 d of exposure, CS CuO/PANI showed higher toxicity to the reproduction parameter, highlighting the importance of a complete study of the NP to better understand its toxicity mechanism.

Preliminary assessment of the performance of oyster shells and chitin materials as adsorbents in the removal of saxitoxin in aqueous solutions.

BACKGROUND: This study evaluated the adsorption capacity of the natural materials chitin and oyster shell powder (OSP) in the removal of saxitoxin (STX) from water. Simplified reactors of adsorption were prepared containing 200 mg of adsorbents and known concentrations of STX in solutions with pH 5.0 or 7.0, and these solutions were incubated at 25°C with an orbital shaker at 200 RPM. The adsorption isotherms were evaluated within 48 hours, with the results indicating a decrease in STX concentrations in different solutions (2-16 µg/L). The kinetics of adsorption was evaluated at different contact times (0-4320 min) with a decrease in STX concentrations (initial concentration of 10 µg/L). The sampling fractions were filtered through a membrane (0.20 µm) and analyzed with high performance liquid chromatography to quantify the STX concentration remaining in solution. RESULTS: Chitin and OSP were found to be efficient adsorbents with a high capacity to remove STX from aqueous solutions within the concentration limits evaluated (> 50% over 18 h). The rate of STX removal for both adsorbents decreased with contact time, which was likely due to the saturation of the adsorbing sites and suggested that the adsorption occurred through ion exchange mechanisms. Our results also indicated that the adsorption equilibrium was influenced by pH and was not favored under acidic conditions. CONCLUSIONS: The results of this study demonstrate the possibility of using these two materials in the treatment of drinking water contaminated with STX. The characteristics of chitin and OSP were consistent with the classical adsorption models of linear and Freundlich isotherms. Kinetic and thermodynamic evaluations revealed that the adsorption process was spontaneous (ΔGads < 0) and favorable and followed pseudo-second-order kinetics.

Induction to oxidative stress by saxitoxin investigated through lipid peroxidation in Neuro 2A cells and Chlamydomonas reinhardtii alga.

Saxitoxin (STX) is a cyanotoxin, which can cause neurotoxic effects and induce ecological changes in aquatic environments, a potential risk to public and environmental health. Many studies of cytotoxicity on animal cells and algae have been performed, although few compare the toxic effects between the two models. In this sense, we investigated the oxidative stress induced by STX (0.4-3.0 nM) in two different cellular models: Neuro-2A (N2A) cells and Chlamydomonas reinhardtii alga by quantification of malondialdehyde (MDA) levels as indicative of lipid peroxidation (LPO). Also was evaluated the antioxidant defense of these cells systems after exposure to STX by the addition of antioxidants in N2A cells culture, and by the measure of antioxidants enzymes activity in C. reinhardtii cells. The MDA levels of N2A cells increased from 15% to 113% for 0.4 and 3.0 nM of STX, respectively, as compared to control. Superoxide-dismutase and catalase did not appear to protect the cell from STX effect while, in cells treated with vitamin E, the rates of MDA production decreased significantly, except for higher concentrations of STX. No MDA productions were observed in algal cells however some effects on antioxidant enzymes activity were observed when algae were exposed to 3.0 nM STX. Our results indicate that the concentrations of STX that may induce oxidative stress through LPO are different in animal and phytoplankton communities. A combination of algal and animal bioassays should be conducted for reliable assessment of oxidative stress induced by STX.