Development of a vibrational startle response assay for screening environmental pollutants and drugs impairing predator avoidance.

The present paper describes the vibrational startle response assay (VSRA), a new robust, simple and automated in vivo medium- to high-throughput procedure for assessment of the escape response and its habituation in zebrafish larvae. Such behaviors enable fish larvae to escape from predator strikes in aquatic ecosystems. The assay is based on measuring the distance moved by each larva during the startle response evoked by repetitive vibrational stimuli. The iterative reduction observed in the response to a series of tapping stimulus in VSRA met the main criteria of habituation. Subsequently, the analysis of concordance using a battery of neuroactive compounds modulating different neurotransmitter systems demonstrated that the results of VSRA are highly predictive of the effects on other vertebrates. Finally, as a proof of concept, VSRA was used to test two relevant environmental pollutants at different concentrations. The results demonstrated that VSRA is suitable for concentration-response analysis of environmental pollutants, opening the possibility to determine the potency and the associated hazard of impaired escape response for the different compounds. Therefore, we suggest that VSRA could be a valuable tool for screening of chemical compounds capable of compromising predator avoidance behavior.

Acrylamide acute neurotoxicity in adult zebrafish.

Acute exposure to acrylamide (ACR), a type-2 alkene, may lead to a ataxia, skeletal muscles weakness and numbness of the extremities in human and laboratory animals. In the present manuscript, ACR acute neurotoxicity has been characterized in adult zebrafish, a vertebrate model increasingly used in human neuropharmacology and toxicology research. At behavioral level, ACR-treated animals exhibited "depression-like" phenotype comorbid with anxiety behavior. At transcriptional level, ACR induced down-regulation of regeneration-associated genes and up-regulation of oligodendrocytes and reactive astrocytes markers, altering also the expression of genes involved in the presynaptic vesicle cycling. ACR induced also significant changes in zebrafish brain proteome and formed adducts with selected cysteine residues of specific proteins, some of them essential for the presynaptic function. Finally, the metabolomics analysis shows a depletion in the monoamine neurotransmitters, consistent with the comorbid depression and anxiety disorder, in the brain of the exposed fish.

Reduction of the Oxidative Stress Status Using Steviol Glycosides in a Fish Model (Cyprinus carpio).

Steviol glycosides are sweetening compounds from the Stevia rebaudiana Bertoni plant. This product is considered safe for human consumption and was approved as a food additive by the Food and Drugs Administration (FDA) and European Food Safety Authority (EFSA). Its effects on the ecosystem have not been studied in depth; therefore, it is necessary to carry out ecotoxicological studies in organisms such as Cyprinus carpio. The present study aimed to evaluate the antioxidant activity by SGs on diverse tissues in C. carpio using oxidative stress (OS) biomarkers. To test the antioxidant activity, carps were exposed to four systems: (1) SGs free control, (2) CCl4 0.5 mL/kg, (3) SGs 1 g/L, and (4) CCl4 0.5 mL/kg + SGs 1 g/L at 96 h. The following biomarkers were analyzed: lipoperoxidation (LPX), hydroperoxide content (HPC), and protein carbonyl content (PCC), as well as antioxidant activity of superoxide dismutase (SOD) and catalase (CAT). It was found that both (3 and 4) systems' exposure decreases LPX, CHP, PCC, SOD, and CAT with respect to the CCl4 system. The results of this study demonstrate that the concentrations of SGs used are not capable of generating oxidative stress and, on the contrary, would appear to induce an antioxidant effect.

Biological hazard evaluation of a pharmaceutical effluent before and after a photo-Fenton treatment.

The aim of this study was to evaluate the biological hazard of a pharmaceutical effluent before and after treatment. For the former, the determined 96h-LC50 value was 1.2%. The photo-Fenton treatment catalyzed with an iron-pillared clay reduced this parameter by 341.7%. Statistically significant increases with respect to the control group (P<0.05) were observed at 12, 24, 48 and 72h in HPC (50.2, 30.4, 66.9 and 43.3%), LPX (22, 83.2, 62.7 and 59.5%) and PCC (14.6, 23.6, 24.4 and 25.6%) and antioxidant enzymes SOD (29.4, 38.5, 32.7 and 49.5%) and CAT (48.4, 50.3, 38.8 and 46.1%) in Hyalella azteca before treatment. Also increases in damage index were observed before treatment of 53.1, 59.9, 66.6 and 72.1% at 12, 24, 48 and 72h, respectively. After treatment the same biomarkers of oxidative stress decreased with respect to before treatment being to HPC (29.3, 22.5, 41.6 and 31.7%); LPX (14.2, 43.1, 30.7 and 35.5%); PCC (12.6, 21.3, 24.2 and 23.9%); SOD (39.2, 33.9, 49.5 and 37.9%) and CAT (28.6, 35.8, 33.7 and 31.7) at 12, 24, 48 and 72h, respectively (P<0.05). The damage index were decreased at 12, 24, 48 and 72h in 48.9, 57.8, 67.3 and 72.1%, respectively. In conclusion, the obtained results demonstrate the need of performing bioassays in order to characterize an effluent before discharge and not base such a decision only upon current normativity. In addition, it was also concluded that the heterogeneous photo-Fenton process decreases the presence of PCT, oxidative stress, genotoxic damage and LC50 in Hyalella azteca.

Oxidative stress induced in Hyalella azteca by an effluent from a NSAID-manufacturing plant in Mexico.

Production in the pharmaceutical industry has increased and along with it, the amount of wastewater of various characteristics and contaminant concentrations. The main chemicals in these effluents are solvents, detergents, disinfectants-such as sodium hypochlorite (NaClO)-and pharmaceutical products, all of which are potentially ecotoxic. Therefore, this study aimed to evaluate the oxidative stress induced in the amphipod Hyalella azteca by the effluent from a nonsteroidal anti-inflammatory drug (NSAID)-manufacturing plant. The median lethal concentration (72 h-LC50) was determined and H. azteca were exposed to the lowest observed adverse effect level (0.0732 %) for 12, 24, 48 and 72 h, and biomarkers of oxidative stress were evaluated [hydroperoxide content (HPC), lipid peroxidation (LPX), protein carbonyl content (PCC), and the activity of the superoxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)]. Statistically significant increases with respect to the control group (P < 0.05) were observed in HPC, LPX and PCC in H. azteca at all exposure times. Antioxidant enzymes activity SOD, CAT and GPx activity also increased significantly (P < 0.05) with respect to the control group. In conclusion, the industrial effluent analyzed in the present study contains NSAIDs and NaClO, and induces oxidative stress in H. azteca.