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.

Oxidative stress and hypermethylation induced by exposure of Oreochromis niloticus to complex environmental mixtures of river water from Cubatão do Sul, Brazil.

In this study, we investigated the effects of oxidative stress and hypermethylation through lipid peroxidation and DNA methylation, respectively, in erythrocytes of Oreochromis niloticus exposed to environmental complex mixture of water from Cubatão do Sul River throughout the year. This river is the source of drinking water for the region of Florianópolis, the capital of Santa Catarina State, Brazil. Lipid peroxidation was quantified by the rate of malondialdehyde (MDA) formation, and DNA methylation was quantified by the rate of 5-methyldeoxycytosine (m(5)dC) formation. In all studied sites, the river water samples caused metabolic changes in O. niloticus. MDA formation rates were significantly different when compared to the negative control (except for samples from Site 1 during spring 2010, summer 2011 and fall 2011). All samples (except Site 1, spring 2010) induced increases in the m(5)dC formation rates, and at the end of the study, the values were near the values found in the positive control (potassium dichromate 2.5mg/L). The results showed that samples of environmental complex mixtures of water from Cubatão do Sul River are capable of inducing high levels of oxidative damage and hypermethylation in O. niloticus.

Toxicity of PAMAM-coated gold nanoparticles in different unicellular models.

Polyamidoamine (PAMAM) dendrimers are used for many pharmaceutical and biomedical applications. However, the toxicological risks of several PAMAM-based compounds are still not fully evaluated, despite evidences of PAMAM deleterious effects on biological membranes, leading to toxicity. In this report, we investigated the toxicity of generation 0 PAMAM-coated gold nanoparticles (AuG0 NPs) in four different models to determine how different cellular systems are affected by PAMAM-coated NPs. Toxicity was evaluated in two mammalian cell lines, Neuro 2A and Vero, in the green alga Chlamydomonas reinhardtii and the bacteria Vibrio fischeri. AuG0 NP treatments reduced cell metabolic activity in algal and bacterial cells, measured by esterase enzymatic activity (C. reinhardtii) and luminescence emission (V. fischeri). EC50 value after 30 min of treatment was similar in both organisms, with 0.114 and 0.167 mg mL(-1) for C. reinhardtii and V. fischeri, respectively. On the other hand, AuG0 NPs induced no change of mitochondrial activity in mammalian cells after 24 h of treatment to up to 0.4 mg mL(-1) AuG0 NPs. Change in the absorption spectra of AuG0 NP in the mammalian cell culture media may indicate an alteration of NP properties that contributed to the low toxicity of AuG0 NPs in mammalian cells. For a safe development of PAMAM-based nanomaterials, the difference of sensitivity between mammalian and microbial cells, as well as the modulation of NPs toxicity by medium properties, should be taken into account when designing PAMAM NPs for applications that may lead to their introduction in the environment.

Effects of exposure to soluble fraction of industrial solid waste on lipid peroxidation and DNA methylation in erythrocytes of Oreochromis niloticus, as assessed by quantification of MDA and m5dC rates.

In this study, lipid peroxidation and DNA methylation were observed in erythrocytes of Oreochromis niloticus exposed to soluble fractions of textile, metal-mechanic and pulp and paper industrial waste, after a period of 48 h. Lipid peroxidation was quantified by the rate of malondialdehyde (MDA) and DNA methylation was quantified by the rate of 5-methyldeoxycytosine (m5dC). Soluble fractions of textile industrial waste caused metabolic changes for all studied samples. In organisms exposed to samples TX1 and TX2 (textile waste) MDA rates were 132.36 and 140.28 nM MDA/mg protein, respectively, while in control organism the MDA rates were 27.5 nM MDA/mg protein. All samples from soluble fractions of textile industrial waste induced increases in m5dC rates, increases varied between 300 percent and 700 percent when compared to the control organism. All the organisms exposed to soluble fractions of metal-mechanic industrial waste presented increases between 360 percent and 600 percent in the rates of MDA, and one of them (sample MM3) induced an increase of 180 percent in the rate of m5dC, when compared to control. Although a significant increase was not observed in the MDA rate of fish exposed to the soluble fractions of pulp and paper industrial waste, there was an increase of 460 percent in the rate of m5dC in one of the samples (sample PP2), when compared to control. The results showed that the soluble fractions of these industrial wastes are capable of inducing oxidative damage and altering the DNA methylation of O. niloticus. Thus, the MDA and m5dC rates demonstrated to be effective biomarkers of exposure, which could be used to evaluate the toxicity of soluble fractions of industrial solid waste.

Investigation of animal and algal bioassays for reliable saxitoxin ecotoxicity and cytotoxicity risk evaluation.

Contamination of water bodies by saxitoxin can result in various toxic effects in aquatic organisms. Saxitoxin contamination has also been shown to be a threat to human health in several reported cases, even resulting in death. In this study, we evaluated the sensitivity of animal (Neuro-2A) and algal (Chlamydomonas reinhardtii) bioassays to saxitoxin effect. Neuro-2A cells were found to be sensitive to saxitoxin, as shown by a 24 h EC50 value of 1.5 nM, which was obtained using a cell viability assay. Conversely, no saxitoxin effect was found in any of the algal biomarkers evaluated, for the concentration range tested (2-128 nM). These results indicate that saxitoxin may induce toxic effects in animal and human populations at concentrations where phytoplankton communities are not affected. Therefore, when evaluating STX risk of toxicity, algal bioassays do not appear to be reliable indicators and should always be conducted in combination with animal bioassays.

Comparative assessment of acute and chronic ecotoxicity of water soluble fractions of diesel and biodiesel on Daphnia magna and Aliivibrio fischeri.

The widespread use of diesel as a transportation fuel and the introduction of biodiesel into the world energy matrix increase the likelihood of aquatic contamination with these fuels. In this case, it is important to know the environmental impacts caused by water-soluble fraction (WSF) of these fuels, since it is the portion that can result in long-term impacts and affect regions far away from the location of a spill. Therefore, we evaluated and compared the aquatic ecotoxicity of the WSF of biodiesel and diesel through acute ecotoxicity tests with the aquatic microcrustacean Daphnia magna and the marine bacteria Aliivibrio fischeri, as well as chronic ecotoxicity tests with D. magna. The WSF of diesel was 2.5-4 folds more toxic than the WSF of biodiesel in acute ecotoxicity tests. Similarly, a comparison of the chronic ecotoxicity demonstrated that the WSF of diesel was more toxic than the WSF of biodiesel. WSF of diesel causes chronic effects on reproduction, longevity and growth of D. magna (NOEC was 12.5, 12.5, 6.25%, respectively), while WSF of biodiesel did not present significantly different results compared to the control for any of the parameters evaluated in any of the dilutions tested (NOEC> 25%). To the best of our knowledge, this is the first study that compares the chronic ecotoxicity of WSF of diesel and biodiesel on D. magna.

Can the surface modification and/or morphology affect the ecotoxicity of zinc oxide nanomaterials?

Among nanomaterials, zinc oxide (ZnO) is notable for its excellent biocidal properties. In particular, it can be incorporated in mortars to prevent biofouling. However, the morphology of these nanomaterials (NMs) and their impact on the action against biofouling are still unknown. This study aimed to assess how the morphology and surface modification can affect the ecotoxicology of ZnO NMs. The morphologies evaluated were nanoparticles (NPs) and nanorods (NRs), and the ZnO NMs were tested pure and with surface modification through amine functionalization (@AF). The toxic effects of these NMs were evaluated by acute and chronic ecotoxicity tests with the well-established model microcrustacean Daphnia magna. The ZnO NMs were characterized by transmission electron microscopy, X-ray diffraction and infrared spectroscopy. The EC5048h to D. magna indicated higher acute toxicity of ZnO@AF NRs compared to all tested NMs. Regarding the chronic test with D. magna, high toxic effects on reproduction and longevity were observed with ZnO@AF NRs and effects on growth were observed with ZnO NRs. In general, all tested ZnO NMs presented high toxicity when compared to the positive control, and the NRs presented higher toxicity than NPs in all tested parameters, regardless of the form tested (pure or with surface modification). Additionally, the pathways of ecotoxicity of the tested ZnO NMs was found to be related to combined factors of Zn ion release, effective diameter of particles and NM internalization in the organism.

Toxicological Evaluation and Quantification of Ingested Metal-Core Nanoplastic by Daphnia magna Through Fluorescence and Inductively Coupled Plasma-Mass Spectrometric Methods.

There are few studies on nanoplastic that propose quantification of the amount ingested combined with evaluation of the toxic effects on aquatic organisms. We propose 2 methods to quantify the amount of polystyrene nanoplastic (PSNP) ingested by Daphnia magna: fluorescence intensity, where a fluorescent monomer (F) is added to the PSNP and quantified through fluorescence light microscopy, and total aluminum quantification, where PSNP is synthesized with Al2 O3 metal-core nanoparticles and used for quantification of the nanoplastic ingested by the organism Daphnia magna using inductively coupled plasma-mass spectrometry. In addition, the PSNP was functionalized with palmitic acid to simulate the environmental conditions leading to biological and chemical transformations. Acute and chronic toxicity tests were performed with fluorescent PSNP (PSNP/F) and palmitic acid-functionalized PSNP/F (PSNP/F-PA). The ingestion quantified was higher by factors of 2.8 and 3.0 for PSNP/F-PA and 1.9 and 1.7 for PSNP/F applying the fluorescence intensity and total Al quantifying methods, respectively, when compared to PSNP. These results are consistent with the data obtained in the toxicity tests, which showed an approximately 3 times increase in the adverse effect of PSNP/F-PA on the mobility and reproduction of the organisms. Thus, the strong inhibition of D. magna reproduction caused by PSNP/F-PA in the chronic toxicity tests could be associated with a greater amount of this nanoplastic being ingested by the organisms. Environ Toxicol Chem 2019;38:2101-2110. © 2019 SETAC.

Effect of chromium oxide (III) nanoparticles on the production of reactive oxygen species and photosystem II activity in the green alga Chlamydomonas reinhardtii.

With the growth of nanotechnology and widespread use of nanomaterials, there is an increasing risk of environmental contamination by nanomaterials. However, the potential implications of such environmental contamination are hard to evaluate since the toxicity of nanomaterials if often not well characterized. The objective of this study was to evaluate the toxicity of a chromium-based nanoparticle, Cr2O3-NP, used in a wide diversity of industrial processes and commercial products, on the unicellular green alga Chlamydomonas reinhardtii. The deleterious impacts of Cr2O3-NP were characterized using cell density measurements, production of reactive oxygen species (ROS), esterase enzymes activity, and photosystem II electron transport as indicators of toxicity. Cr2O3-NP exposure inhibited culture growth and significantly lowered cellular Chlorophyll a content. From cell density measurements, EC50 values of 2.05±0.20 and 1.35±0.06gL(-1) Cr2O3-NP were obtained after 24 and 72h of exposure, respectively. In addition, ROS levels were increased to 160.24±2.47% and 59.91±0.15% of the control value after 24 and 72h of exposition to 10gL(-1) Cr2O3-NP. At 24h of exposure, the esterase activity increased to 160.24% of control value, revealing a modification of the short-term metabolic response of algae to Cr2O3-NP exposure. In conclusion, the metabolism of C. reinhardtii was the most sensitive to Cr2O3-NP after 24h of treatment.

Comparative evaluation of acute and chronic toxicities of CuO nanoparticles and bulk using Daphnia magna and Vibrio fischeri.

Copper oxide (CuO) has various applications, as highlighted by the incorporation of this compound as a biocide of antifouling paints for coating ships and offshore oil platforms. The objective of this study was to evaluate and compare the aquatic toxicity of CuO nanoparticles (NPs) and microparticles (MPs) through acute and chronic toxicity tests with the freshwater microcrustacean Daphnia magna and an acute toxicity test with the bioluminescent marine bacteria Vibrio fischeri. Acute toxicity results for D. magna in tests with CuO NPs (EC50, 48 h=22 mg L(-1)) were ten times higher than those for tests with CuO MPs (EC50, 48 h=223.6 mg L(-1)). In both periods of exposure of V. fischeri, the CuO NPs (EC50, 15m 248±56.39 - equivalent to 12.40%; EC50, 30 m 257.6±30.8 mg L(-1) - equivalent to 12.88%) were more toxic than the CuO MPs (EC50, 15m 2404.6±277.4 - equivalent to 60.10%; EC50, 30 m 1472.9±244.7 mg L(-1) - equivalent to 36.82%). In chronic toxicity tests, both forms of CuO showed significant effects (p<0.05) on the growth and reproduction parameters of the D. magna relative to the control. Additionally, morphological changes, such as lack of apical spine development and malformed carapaces in D. magna, were observed for organisms after the chronic test. The toxicity results demonstrate that CuO NPs have a higher level of toxicity than CuO MPs, emphasizing the need for comparative toxicological studies to correctly classify these two forms of CuO with identical CAS registration numbers.

Evaluation of toxicity and oxidative stress induced by copper oxide nanoparticles in the green alga Chlamydomonas reinhardtii.

Copper oxide nanoparticles (CuO NP) are frequently employed for their antimicrobial properties in antifouling paints. Their extensive use can contaminate aquatic ecosystems. However, the toxicological effects of this NP in the environment are poorly known. In this study, we evaluated the toxicity and oxidative stress induced by CuO NP on Chlamydomonas reinhardtii using several toxicological assays. CuO NP was found to induce growth inhibition and a significant decrease in carotenoids levels. From data on cells density after 72 h of CuO NP exposure in light, the EC50 value was calculated to be 150.45±1.17 mg L(-1) and the NOEC≤100 mg L(-1). Evaluation of esterase activity demonstrates a decrease in cell metabolism activity with the increase of CuO NP concentration. The CuO NP induced an increase of reactive species level (190±0.45% at 1000 mg L(-1) after 72 h of exposition, compared to control) and lipid peroxidation of cellular membranes (73±2% at 1000 mg L(-1) of CuO NP in 72 h of exposition, compared to control). Investigation of CuO NP uptake showed the presence of NP into C. reinhardtii cells in different sites of the cell and, biomarkers of enzymatic antioxidants showed a change of activity after CuO NP exposition. In conclusion, C. reinhardtii was shown to be sensitive to the presence of CuO NP in their environment and CuO NP treatments induced a toxic response from 0.1 mg L(-1) after 72 h of treatment.

Polymer coating of copper oxide nanoparticles increases nanoparticles uptake and toxicity in the green alga Chlamydomonas reinhardtii.

Copper oxide nanoparticles (CuO NPs) are frequently used in a polymer-coated form, to be included in paints or fabrics for antimicrobial properties. Their application in antifouling paints may lead to the contamination of aquatic ecosystems. However, the toxicological risk of NPs in the environment is hard to evaluate due to a lack of knowledge on the mechanisms of NP interaction with biological systems. In this study, we investigated the effect of polymer coating on CuO NP toxicity in the green alga Chlamydomonas reinhardtii by comparing bare and polymer-coated CuO NPs prepared from the same CuO nanopowder. Both CuO NP suspensions were toxic to C. reinhardtii after 6 h treatment to concentrations of 0.005-0.04 g L(-1). Bare and polymer-coated CuO NPs induced a decrease of Photosystem II activity and the formation of reactive oxygen species. Polymer-coated CuO NP was found to be more toxic than the uncoated CuO NP. The higher toxicity of CS-CuO NP was mainly associated with the increased capacity of polymer-coated CuO NP to penetrate the cell compared to bare CuO NPs. These results indicates that the high toxicity of polymer-coated CuO NPs in algal cells results of intracellular interactions between NPs and the cellular system.