Neuromotor activity inhibition in zebrafish early-life stages after exposure to environmental relevant concentrations of caffeine.

Caffeine (CAF), a neuroactive compound, has been found in surface waters at concentrations ranging from few nanograms up to micrograms and may induce adverse effects in aquatic vertebrates. Thus, the aim of this study was to evaluate the potential of CAF in affecting fish early-life stages in a wide concentration range, including occurring levels in surface waters. Specimens of zebrafish in early-life stages were exposed to CAF for 168 h and survival, developmental alterations, locomotor activity and acetylcholinesterase activity were evaluated. CAF induced mortality in embryos unable to hatch or in larvae after hatching (LC50 - 168 h = 283.2 mg/L). Tail deformities were observed in organisms exposed to concentrations ≥ 40 mg/L, while edemas were found at concentrations of 100 mg/L. CAF also decreased the total swimming time and distance moved of exposed organisms (LOEC = 0.0006 mg/L). Locomotor inhibition may be associated with an acetylcholinesterase inhibition observed at concentration ≥ 0.0088 mg/L. Therefore, the hazard of CAF for fish populations deserves further attention since unexpected effects on neuro-behavioral parameters occurs at concentrations often detected in natural aquatic ecosystems.

Auramine dyes induce toxic effects to aquatic organisms from different trophic levels: an application of predicted non-effect concentration (PNEC).

The dyes Auramine and Auramine O are used in several industrial products, despite the scarce information regarding their ecotoxicity. The aim of the present study was to assess the acute and chronic toxicity of both dyes to aquatic organisms from different trophic levels (Raphidocelis subcapitata, Daphnia similis, Hydra attenuata, and Danio rerio) and calculate their predicted non-effect concentrations (PNEC). Auramine and Auramine O induced toxicity to all selected test organisms with L(E)C50 values ranging from 300 to 4800 ug/L. Both dyes induced inhibition in the growth rate of exposed algae, negatively affecting the reproduction of D. similis and induced deformities in H. attenuata (clubbed tentacles and shortened tentacles) and D. rerio (edemas, tail malformation and delay in yolk sac absorption). PNEC values of 0.92 µg/L and 4.0 µg/L were obtained for Auramine and Auramine O, respectively, based on results of the most sensitive test system (algae). Test results were analyzed using the Criteria of Reporting and Evaluating Ecotoxicity Data (CRED), confirming their reliability and relevance. Thus, PNEC values can be used in future risk assessments of those substances in freshwater systems.

Cytotoxic and genotoxic effects of two hair dyes used in the formulation of black color.

According to the International Agency for Research on Cancer (IARC), some hair dyes are considered mutagenic and carcinogenic in in vitro assays and exposed human populations. Epidemiological studies indicate that hairdressers occupationally exposed to hair dyes have a higher risk of developing bladder cancer. In Brazil, 26% of the adults use hair dye. In this study, we investigated the toxic effects of two hair dyes, Basic Red 51 (BR51) and Basic Brown 17 (BB17), which are temporary dyes of the azo group (R-N=N-R'), used in the composition of the black hair dye. To this end, MTT and trypan blue assays (cytotoxicity), comet and micronucleus assay (genotoxicity) were applied, with HepG2 cells. For cytotoxic assessment, dyes were tested in serial dilutions, being the highest concentrations those used in the commercial formula for hair dyes. For genotoxic assessment concentrations were selected according to cell viability. Results showed that both dyes induced significant cytotoxic and genotoxic effects in the cells, in concentrations much lower than those used in the commercial formula. Genotoxic effects could be related to the azo structure present in the composition of the dyes, which is known as mutagenic and carcinogenic. These results point to the hazard of the hair dye exposure to human health.

Assessment of cytotoxicity and AhR-mediated toxicity in tropical fresh water sediments under the influence of an oil refinery.

Oil refinery effluents contain many chemicals at variable concentrations. Therefore, it is difficult to predict potential effects on the environment. The Atibaia River (SP, Brazil), which serves as a source of water supply for many municipalities, receives the effluents of one of the biggest oil refinery of this country. The aim of this study was to identify the (eco)toxicity of fresh water sediments under the influence of this oil refinery through neutral red (cytotoxicity) and ethoxyresorufin-O-deethylase (EROD) assays (AhR-mediated toxicity) in RTL-W1 cells (derived from fish liver). Once the refinery captures the waters of Jaguarí River for the development of its activities and discharges its effluents after treatment into the Atibaia River, which then flows into Piracicaba River, sediments from both river systems were also investigated. The samples showed a high cytotoxic potential, even when compared to well-known pollution sites. However, the cytotoxicity of samples collected downstream the effluent was not higher than that of sediments collected upstream, which suggested that the refinery discharges are not the main source of pollution in those areas. No EROD activity could be recorded, which could be confirmed by chemical analyses of polycyclic aromatic hydrocarbons (PAHs) that revealed a high concentration of phenanthrene, anthracene, fluoranthene, and pyrene, which are not EROD inducers in RTL-W1 cells. In contrast, high concentrations of PAHs were found upstream the refinery effluent, corroborating cytotoxicity results from the neutral red assay. A decrease of PAHs was recorded from upstream to downstream the refinery effluent, probably due to dilution of compounds following water discharges. On the other hand, these discharges apparently contribute specifically to the amount of anthracene in the river, since an increase of anthracene concentrations could be recorded downstream the effluent. Since the extrapolation of results from acute toxicity to specific toxic effects with different modes of action is a complex task, complementary bioassays covering additional specific effects should be applied in future studies for better understanding of the overall ecotoxicity of those environments.

Changes in toxicity and dioxin-like activity of sediments from the Tietê River (São Paulo, Brazil).

Acute cytotoxicity (neutral red assay), AhR-mediated toxicity (CYP P450 1A induction potential-EROD assay) and concentrations of PAHs were recorded in RTL-W1 cells to assess the ecotoxicological potential of sediments from reservoirs along the Tietê River and the Pinheiros River (Brazil). Almost all sediments tested induced cytotoxicity and stimulated cytochrome P450-associated EROD activity. Toxicity increases from Tietê River spring to São Paulo city region and decreases towards downstream. A closer analysis of chemical measurements of PAHs and results from bioassay revealed that the PAHs analyzed could not explain more than 7% of the EROD-inducing potencies. Results confirm that most of the toxicity is due to the discharges of the metropolitan area of São Paulo. Moreover, they indicate additional sources of pollutants along the river course, which contribute to the degradation of each reservoir.