Toxicity of dyes to zebrafish at the biochemical level: Cellular energy allocation and neurotoxicity

doi:10.1016/j.envpol.2017.12.020

Environmental Pollution

Volume 235, April 2018, Pages 255-262

https://doi.org/10.1016/j.envpol.2017.12.020 Get rights and content

Highlights

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Zebrafish embryos consumed less energy, compromising their energy balance.
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Energy allocation impairment reduced compromised locomotor activity of zebrafish larvae.
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Effects on the cholinergic system were not necessary for locomotor impairment.
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After photodegradation, the natural dye completely lost its toxicity.

Abstract

Dyes are widely distributed worldwide, and can be found in wastewaters resulting from industrial or urban effluents. Dyes are of particular concern as contaminants of the aquatic environment, since their toxicity remain poorly understood. Thus, the current study was designed to assess the effects induced by the synthetic azo dye Basic Red 51 (BR51) and by the natural naphthoquinone dye erythrostominone (ERY) on zebrafish early life stages (Danio rerio) at different biological organization levels, i.e., studying how changes in biochemical parameters of important physiological functions (neurotransmission and cellular energy allocation) may be associated with behavior alterations (swimming activity). This approach was also used to assess the effects of ERY after its photodegradation resulting in a colorless product(s) (DERY). Results showed that after 96 h exposure to BR51 and Ery, zebrafish embryos consumed less energy (LOEC = 7.5 mg/L), despite the unaltered levels of available energy (carbohydrates, lipids and proteins). Hence, cellular energy allocation (CEA) was significantly increased. On the other hand, only ERY decreased the acetylcholinesterase activity (LOEC = 15 mg/L). Despite that, zebrafish larvae exposed to both dyes until 144 h were less active. In contrast, DERY did not affect any parameter measured. These results indicate an association between a decrease consumption of energy and decrease swimming activity resulting from an environmental stress condition, independently of the neurotoxicity of the dyes. Degradation of ERY by light prevented all toxic effects previously observed, suggesting a cheap, fast and easy alternative treatment of effluents containing this natural dye. All tools assessed in our current study were sensitive as early-warning endpoints of dyes toxicity on zebrafish early life stages, and suggest that the CEA assay might be useful to predict effects on locomotor activity when cholinergic damage is absent.

Graphical abstract

Section snippets

Introdution

Synthetic dyes containing the azo-aromatic chromophore group comprise the largest chemical class of dyes used worldwide (Guaratini and Zanoni, 2000), but these are relatively persistent pollutants due to their high stability (O'Neill et al., 1999, Chung, 2000), hindering the removal or reduction of toxicity of dye-containing industrial wastewaters by conventional treatments. It is known that textile industries release approximately 15–50% of the total amount of dyes used during manufacturing

Chemicals

Basic Red 51 (BR51) (2-(((4-dimethyllamino) phenyl) azo)-1,3-dimethyl-1H-imidazolium chloride, CAS No. 77061-58-6, MW: 279.6 g/mol, purity 99% a.i.) (Fig. 1a) was purchased from LCW Import and Export Ltd (São Paulo, Brazil). BR51 stock solutions were directly prepared in reverse osmosis water at 100 mg/L final concentration. The endophytic fungus LC01-A strain (in identification phase) was kindly provided from Coleção de Microrganismos de Importância Agrícola e Ambiental (CCMA, EMBRAPA,

Energy available, energy consumed and cellular energy allocation

Lipids were the main source of energy reserves of zebrafish embryos (88–90%), followed by carbohydrates (6–9%) and proteins (2.5–4.5%). BR51 (F_4;25 = 0.71, p > .05), ERY (F_4;29 = 1.44, p > .05) and DERY (F_4;25 = 0.49, p > .05) did not significantly change Ea, compared to control group (Table 1).

However, zebrafish embryos exposed to both BR51 and ERY consumed significantly less energy at concentrations equal or above 7.5 mg/L (F_4;29 = 3.96, p < .05 and F_4;29 = 23.72, p < .05, respectively),

Discussion

Cellular energy allocation results showed that lipids were the major energy available (around 89%) on zebrafish embryos, followed by carbohydrates (around 7.5%) and proteins (around 3.5%). Indeed, lipids represent the first energy source mobilized when organisms are exposed to chemicals, as reported for Daphnia magna (De Coen and Janssen, 1997), and along with carbohydrates, they are quickly mobilized to supply a sudden energy demand (Smolders et al., 2003). On the other hand, proteins were the

Conclusions

Our results corroborate the usefulness of energy budget biomarkers as an important tool to understand how organisms under stress use their energy reserves. Moreover, those parameters describing Ec and CEA were associated with the impairment of the locomotor activity of zebrafish early life stages exposed to dyes: both natural (ERY) and synthetic (BR51) dyes decrease the Ec of zebrafish embryos, compromising the energy balance and hence the energy available for allocation to other biological

Author contributions

All authors contributed equally.

Conflicts of interest

The authors declare no conflicting interests.

Acknowledgment

We thank the financial support of São Paulo Research Foundation (FAPESP Processes No. 2013/14397-0, 2014/27009-0 and 2013/01509-4, São Paulo, Brazil), and Fundação para a Ciência e a Tecnologia (FCT) and POPH/FSE (Programa Operacional Potencial Humano/Fundo Social Europeu) for the research contract of C. Gravato (IF/01401/2014).

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Toxicity of dyes to zebrafish at the biochemical level: Cellular energy allocation and neurotoxicity☆

Highlights

Abstract

Graphical abstract

Section snippets

Introdution

Chemicals

Energy available, energy consumed and cellular energy allocation

Discussion

Conclusions

Author contributions

Conflicts of interest

Acknowledgment

Chemosphere

Aquat. Toxicol.

Dyes Pigm

J. Clean. Prod.

Food Chem. Toxicol.

Toxicol. Vitro

Int. J. Pharm.

Food Chem.

Chemosphere

J. Environ. Chem. Eng

Cell Rep

Aquat. Toxicol.

Comp. Biochem. Physiol. C

Phytochemistry

Ocean Coast. Manag.

J. Fluorine Chem.

Environ. Pollut. A

Ecotoxicol. Environ. Saf.

LWT

Ind. Crop. Prod.

Ecotoxicol. Environ. Saf.

Sci. Total Environ.

J. Clean. Prod.

Environ. Pollut.

Mar. Environ. Res.

Reprod. Toxicol.

Food Chem. Toxicol.

Environ. Int.

Carbohyd. Polym

J. Exp. Mar. Biol. Ecol.

Toxicol. Lett.

J. Hazard Mater.