Aplicação dos princípios do RISK21 na avaliação do risco de inseticidas da classe dos piretróides / The use of RISK21 principles in the risk assessment of pyrethroids

A avaliação do risco é um processo científico e sistemático que incorpora quantitativamente o perigo e a exposição a diversos agentes. O processo de avaliação do risco tem evoluído nos últimos anos, indo além da exposição a únicos agentes e vias de exposição para a caracterização do risco cumulativo a múltiplos agentes. As metodologias para avaliação do risco cumulativo não são harmonizadas o que pode tornar o processo complexo. Nesta linha, a abordagem do RISK21 promovida pelo Health Environmental Science Institute (HESI) pode contribuir para desmistificar o tema. A exposição combinada da ingestão de resíduos de praguicidas através da dieta e do uso residencial de produtos a base de piretróides pela população brasileira não são conhecidas. Os piretróides são praguicidas utilizados na lavoura, bem como em ambiente doméstico no controle de pragas. O mecanismo de toxicidade destes agentes é bem conhecido e de relevância para a saúde humana, pois atuam sobre a permeabilidade iônica dos canais de sódio sensíveis a voltagem (CSSV), produzindo efeitos na excitabilidade das terminações nervosas. Como os seres humanos são potencialmente expostos a estes agentes, portanto, torna-se importante compreender os riscos cumulativos da exposição a estes praguicidas pela população brasileira. O objetivo deste trabalho foi conduzir a avaliação do risco dos piretróides registrados no Brasil com base nos princípios do RISK21. A abordagem em etapas proposta pelo RISK21 demonstrou que o risco da ingestão crônica e aguda de resíduos de piretróides foi considerado aceitável. Além disso, não foi observada qualquer preocupação toxicológica decorrente da exposição residencial a estes agentes. Quando combinados os cenários da dieta aguda e residencial, também não foram observados níveis de preocupação, portanto, o risco foi considerado aceitável. A avaliação do risco dos piretróides registrados para o uso agrícola e residencial no Brasil com base nos principios do RISK21 foi uma importante etapa neste trabalho, uma vez que foi possível avaliar o risco e preocupações para cada um dos praguicidas de maneira rápida e visual. Além disso, mesmo considerando premissas altamente conservadoras, observou-se que a população exposta de maneira combinada a estes agentes não demonstrou um nível de preocupação para o cenário brasileiro

Risk assessment is a scientific and systematic approach that quantitatively incorporates hazard and exposure to agents' evaluation. The risk assessment process has evolved in recent years, going beyond exposure to single agents and pathways to characterize multiple agents' cumulative risk. Cumulative risk assessment methodologies are not harmonized, which can make the process complex. In this line, the RISK21 approach promoted by the Health Environmental Science Institute (HESI) can demystify the subject. The combined exposure of residue intake through diet and residential use of pyrethroid-based products by the Brazilian population is unknown. Pyrethroids are pesticides used in the crop as well as in a domestic environment in pest control. The mechanism of toxicity of these agents is well known and relevant to human health, as they act on the ionic permeability of voltage-sensitive sodium channels (VSSC), producing effects on the excitability of nerve endings. As human beings are potentially exposed to these agents, it is essential to understand the cumulative risks derived from the exposure to these pesticides by the Brazilian population. The objective of this research was to conduct the risk assessment based on the principles of RISK21 of pyrethroids registered in Brazil. The stepwise approach proposed by RISK21 demonstrated that the risk of chronic and acute ingestion of pyrethroid residues was considered acceptable. Furthermore, no toxicological concern stemming from residential exposure to these agents was observed. When acute and residential diet scenarios were combined, no levels of concern were also observed, so the risk was considered acceptable. The risk assessment based on the principles of RISK21 of pyrethroids registered for agricultural and residential use in Brazil was an essential step in this research since it was possible to assess the risk and concerns for each of the pesticides in a fast and visual way. Moreover, from highly conservative premises, it was observed that the population exposed in combination with these agents did not demonstrate a level of concern for the Brazilian scena

Antioxidant capacity of omega-3-fatty acids and vitamin E against imidacloprid-induced hepatotoxicity in Japanese quails.

Imidacloprid (IM) is a neonicotinoid insecticide, used in a wide range of agricultural activities worldwide. However, it results in ecosystem disturbances and signs of toxicity in human and animals. The current study was designed to elucidate the protective effects of omega-3-fatty acids (OFAs) and vitamin E (Vit E) against IM hepatotoxicity in Japanese quails. Seventy male quails (30 days old) were divided into seven groups (n = 10); G1 -ve control; G2 received IM (+ve control); G3 received OFA; G4 received Vit E; and G5, G6, and G7 received OFA and/or Vit E with IM for 30 days, respectively. Blood and liver tissue samples were collected. Imidacloprid significantly (p < 0.05) increased serum levels of alanine transferase (ALT), aspartate transferase (AST), triglycerides (TGC), and low-density lipoprotein cholesterol (LDL-C), as well as liver tissue malondialdehyde (MDA) concentration. Moreover, IM caused a significant (p < 0.05) decrease in the levels of serum high-density lipoprotein cholesterol (HDL-C), as well as liver superoxide dismutase (SOD) enzyme activity and reduced-glutathione (GSH) concentration in comparison to the -ve control group. Histopathological changes in hepatocytes, including thick cell trabeculae with marked hydropic vacuolar degeneration of cytoplasm, were found in IM-treated group. Treatment with OFA and/or Vit E resulted in significant improvements in general body condition, serum HDL-C level, and liver tissue SOD enzyme activity and GSH concentration, as well as significant decreases in the levels of serum AST, ALT, TGC, LDL-C, and hepatic tissue MDA. In conclusion, OFA and Vit E have a protective effect against IM toxicity, especially in their combination.

Species specific RNA A-to-I editing of mosquito RDL modulates GABA potency and influences agonistic, potentiating and antagonistic actions of ivermectin.

The insect GABA receptor, RDL, is the target of several classes of pesticides. The peptide sequences of RDL are generally highly conserved between diverse insects. However, RNA A-to-I editing can effectively alter amino acid residues of RDL in a species specific manner, which can affect the potency of GABA and possibly insecticides. We report here that RNA A-to-I editing alters the gene products of Rdl in three mosquito disease vectors, recoding five amino acid residues in RDL of Aedes aegypti and six residues in RDLs of Anopheles gambiae and Culex pipiens, which is the highest extent of editing in RDL observed to date. Analysis of An. gambiae Rdl cDNA sequences identified 24 editing isoforms demonstrating a considerable increase in gene product diversity. RNA editing influenced the potency of the neurotransmitter, GABA, on An. gambiae RDL editing isoforms expressed in Xenopus laevis oocytes, as demonstrated by EC50s ranging from 5 ± 1 to 246 ± 41 µM. Fipronil showed similar potency on different editing isoforms, with IC50s ranging from 0.18 ± 0.08 to 0.43 ± 0.09 µM. In contrast, editing of An. gambiae RDL affected the activating, potentiating and inhibiting actions of ivermectin. For example, ivermectin potentiated currents induced by GABA at the EC20 concentration in the unedited isoform but not in the fully edited variant. Editing of a residue in the first transmembrane domain or the cys-loop influenced this potentiation, highlighting residues involved in the allosteric mechanisms of cys-loop ligand-gated ion channels. Understanding the interactions of ivermectin with molecular targets may have relevance to mosquito control in areas where people are administered with ivermectin to treat parasitic diseases.

PPAR-γ activation attenuates deltamethrin-induced apoptosis by regulating cytosolic PINK1 and inhibiting mitochondrial dysfunction.

Central events in the mitochondrial-dependent cell death pathway include the disruption of mitochondrial membrane potential, which causes the release of apoptogenic molecules leading to cell death. Based on the cytotoxic mechanism of deltamethrin (DLM), we examined the neuroprotective mechanisms of rosiglitazone (RGZ), which is against DLM-induced neuronal cell death. In this study, we found that DLM induces apoptosis in SH-SY5Y cells as demonstrated by the activation of caspase-3 and nuclear condensation. In addition, neuronal cell death in response to DLM was due to mitochondrial dependent-apoptosis pathways since DLM increased cytochrome c release into the cytosol and activated caspase-9. DLM exposure reduced PINK1 expression, and pretreatment with RGZ significantly reduced cytochrome c release and caspase-9 activation. RGZ also attenuated the reduction of complex I activity, mitochondrial membrane potential, and ATP levels. Pretreatment with RGZ significantly enhanced PINK1 expression in DLM-exposed cells. In addition, RGZ increased cytosolic PINK1 by inhibiting mitochondrial translocation of PINK1. Interestingly, RGZ fails to rescue DLM-induced mitochondrial dysfunction both in PINK1 knockdown and PPAR-γ antagonist treated cells. Results from this study suggest that RGZ exerts anti-apoptotic effects against DLM-induced cytotoxicity by attenuation of mitochondrial dysfunction through cytosolic PINK1-dependent signaling pathways.

In vitro effect of N-acetylcysteine on hepatocyte injury caused by dichlorodiphenyltrichloroethane and its metabolites.

The organochlorine pesticide, dichlorodiphenyltrichloroethane (DDT), is still used to combat the spread of malaria in several developing countries despite its accumulation and known hepatotoxic effects that have been demonstrated both in vitro and in vivo. N-Acetylcysteine (NAC) is a recognized hepatoprotective agent that has been reported to reduce hepatotoxicity initiated by many different compounds. The aim of this study was to determine whether NAC could counter in vitro hepatocyte injury induced by DDT or its two major metabolites, dichlorodiphenyldichloroethylene and dichlorodiphenyldichloroethane. HepG2 cell cultures were used to assess the following parameters of toxicity: cellular viability, intracellular levels of reactive oxygen species (ROS), mitochondrial membrane potential and initiation of apoptosis. None of the three test compounds induced ROS generation, yet exposure to any of the three compounds produced mitochondrial hyperpolarization, which was countered by NAC pretreatment. All three test compounds also induced apoptotic cell death, which was inhibited by NAC. Despite NAC counteracting some adverse intracellular changes due to organochlorine exposure, it appeared to aggravate the cytotoxic effects of the organochlorine compounds at low test concentrations. As the same outcome may also occur in vivo, results from the present study raise concern about the use of NAC as treatment for DDT-induced hepatotoxicity.

Comparison of the activity of selamectin, imidacloprid and fipronil for the treatment of dogs infested experimentally with Ctenocephalides canis and Ctenocephalides felis felis.

Twenty-four beagles were randomly allocated into four groups of six and housed in separate cages. Each dog was infested with 25 Ctenocephalides canis and 25 Ctenocephalides felis felis and two days later (day 0) the dogs in groups 1, 2 and 3 received a spot-on application of selamectin (6 mg/kg), imidacloprid (10 mg/kg), or fipronil (6-7 mg/kg), respectively, while the dogs in group 4 were not treated. The dogs were combed 48 hours later, the fleas were removed, counted and their species were determined. All the dogs were reinfested with the same number of the two species of fleas on days 7, 14, 21, 28 and 35, and the efficacy of the treatments was calculated 48 hours after each infestation. The mean numbers of fleas on the control dogs were 19.8 C. canis and 14.7 C. felis felis. The three treatments were effective for the full 35 days of the trial; over the first 28 days, the efficacy of selamectin ranged from 81 to 100 and 92 to 99 per cent against C. felis felis and C canis, respectively, the efficacy of imidacloprid ranged from 98 to 100 per cent and the efficacy of fipronil was 100 per cent against both species. There were no significant differences between the three treatments.