Glyphosate (N-phosphonomethyl-glycine) (GLY) is the active ingredient of the most used herbicides in the world. GLY is applied in formulated products known as glyphosate-based herbicides (GBH), which could induce effects that are not predicted by toxicity assays with pure GLY. This herbicide is classified as organophosphorus compound, which is known to induce neurotoxic effects. Although this compound is classified as non-neurotoxic by regulatory agencies, acute exposure to GBH causes neurological symptoms in humans. However, there is no consensus in relation to neurotoxic effects of GBH. Thus, the aim of this study was to investigate the neurotoxic effects of the GBH in the zebrafish Danio rerio, focusing on acute toxicity, the activity and transcript levels of mitochondrial respiratory chain complexes, mitochondrial membrane potential, reactive species (RS) formation, and behavioral repertoire. Adult zebrafish were exposed in vivo to three concentrations of GBH Scout, which contained GLY in formulation (fGLY) (0.065, 1.0 and 10.0 mg L fGLY) for 7 d, and an in vitro assay was performed using also pure GLY. Our results show that GBH induced in zebrafish brain a decrease in cell viability, inhibited mitochondrial complex enzymatic activity, modulated gene expression related to mitochondrial complexes, induced an increase in RS production, promoted hyperpolarization of mitochondrial membrane, and induced behavioral impairments. Together, our data contributes to the knowledge of the neurotoxic effects of GBH. Mitochondrial dysfunction has been recognized as a relevant cellular response that should not be disregarded. Moreover, this study pointed to the mitochondria as an important target of GBH.
Roundup is a glyphosate-based herbicide (GBH) widely used in agriculture and may cause toxic effects in non-target organisms. Model organisms, as zebrafish, and analysis of gene expression by reverse transcription-quantitative PCR (RT-qPCR) could be used to better understand the Roundup toxicity. A prerequisite for RT-qPCR is the availability of appropriate reference genes; however, they have not been described for Roundup-exposed fish. The aim of this study was to evaluate the expression stability of six reference genes (rpl8, β-act, gapdh, b2m, ef1α, hprt1) and one expressed repetitive element (hatn10) in organs of males (brain, gill, testis) and females (ovary) of zebrafish exposed to Roundup WG at three concentrations (0.065, 0.65 and 6.5 mg N-(phosphonomethyl) glycine/l) for 7 days. Genes were ranked by geNorm, NormFinder, BestKeeper, Delta C t and RefFinder, and their best combinations were determined by geNorm and NormFinder programs. The two most stable ranked genes were specific to each organ: gill (β-act; rpl8); brain (rpl8; β-act); testis (ef1α; gapdh); and ovary (rpl8; hprt1). The cat transcript level was used to evaluate the effect of normalization with these reference genes. These are the first suitable reference genes described for the analysis of gene expression in organs of Roundup-exposed zebrafish, and will allow investigations of the molecular mechanisms of Roundup toxicity.
The use of toxic pesticides has become a world problem because they can contaminate streams and rivers, producing an adverse impact on non-target aquatic biota, including fishes. Cypermethrin is one of the most important insecticides to control ectoparasites in wide-scale. The aim of this study was to evaluate the effect of commercial formulations of cypermethrin, SHERPA O (0.0, 0.075, 0.15, and 0.3 µg/L of cypermethrin) in fish Prochilodus lineatus for 96 h in semi-static condition, using biomarkers of genotoxicity: micronucleus frequency (MNF) in erythrocytes and biomarkers of oxidative damage: lipid peroxidation (TBARS) and antioxidant defenses, catalase (CAT) and glutathione (GSH) in liver tissue. Our results showed a significant decrease (p < 0.05) of CAT at pesticide concentrations of 0.150 and 0.300 μg/L, but no significant difference was observed in TBARS or GSH in any exposed group (p > 0.05) in comparison to the control. A significant increase was observed in the MNF in the group exposed to 0.3 μg/L of cypermethrin compared to negative control (p < 0.05). Finally, P. lineatus proved to be a sensitive species to the commercial formulations of cypermethrin and that CAT and MNF are effective indicators of these toxic effects.
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