Análise dos efeitos do glifosato e sua formulação Roundup® nas células e gônadas dos peixes Danio rerio (Cyprinidae) / Analysis of the effects of glyphosate and its formulation Roundup® on the cells and gonads of fish Danio rerio (Cyprinidae)

Os herbicidas estão entre os insumos químicos mais utilizados na agricultura e apresentam ação toxicológica para as plantas. Dentre os herbicidas existentes, o glifosato e a sua formulação comercial Roundup® são os mais utilizados devido à eficácia no controle de ervas daninhas. Entretanto, estudos sugerem que estes herbicidas podem provocar distúrbios a organismos aquáticos, principalmente o Roundup®, que além do glifosato ativo, apresenta na composição componentes surfactantes que aumentariam o seu potencial toxicológico para os seres vivos. Desta forma, o presente estudo teve como objetivo comparar o potencial mutagênico e desenvolvimento gonadal do princípio ativo glifosato e sua formulação Roundup® em peixes Danio rerio. Para isso, exemplares adultos de D. rerio permaneceram em aquários expostos à concentração de 65 µg/L de cada herbicida. Foram realizadas contagens de micronúcleos, calculado o índice gonadossomático (IGS) e verificado os estágios de maturação gonadal. Os resultados demonstraram que os peixes expostos aos herbicidas apresentaram aumentos significativos na frequência de micronúcleos, bem como no IGS, e maior ocorrência de ovários com estágios em maturação e maduros. Não foram verificadas diferenças significativas na comparação dos herbicidas, exceto um aumento na frequência de ovários maduros nas fêmeas expostas ao glifosato. Observou-se que, mesmo em pequenas concentrações, os herbicidas poderiam ter ocasionado efeitos nocivos para os peixes D. rerio, podendo induzir efeitos clastogênicos nos eritrócitos, assim como distúrbios reprodutivos, aumentando a preocupação sobre os impactos que estes herbicidas podem acarretar sobre o ciclo de vida destes organismos.(AU)

The herbicides are among the most used chemical inputs in the agriculture and present a great toxicological action for the plants. Among the existing herbicides, glyphosate and its commercial formulation Roundup® are the most used due to their effectiveness in control of weeds. However, studies suggest that these herbicides can cause disturbances to aquatic organisms, mainly the Roundup®, that besides active glyphosate, it presents in the composition components surfactants that would increase its toxicological potential for the alive beings. Therefore, the aim of this study is to compare the mutagenic potential and gonadal development of the active principle glyphosate and its formulation Roundup® in fish Danio rerio. Thus, adult copies of D. rerio stayed in aquariums exposed to a concentration of 65 µg / L of each herbicide. Micronucleus counts were performed and the Gonadossomatic index (GSI) was calculated and the stages of gonadal maturation were verified. The results demonstrated that the fish exposed to the herbicides, presented significant increases in the micronucleus frequency, as well as in GSI and larger occurrence of ovaries with stages in maturation and mature. There were no significant differences in herbicide comparison, except an increase in the frequency of mature ovaries in the exposed females to glyphosate. It was observed that even in small concentrations, the herbicides could presented harmful effects for the fish D. rerio, could have caused clastogenic effects in the erythrocytes, as well as, reproductive disturbances, increasing the concern about the impacts that these herbicides can have on the life cycle of these organisms.(AU)

Identification of endogenous normalizing genes for expression studies in inguinal ring tissue for scrotal hernias in pigs.

The use of reference genes is required for relative quantification in gene expression analysis and since the stability of these genes could be variable depending on the experimental design, it has become indispensable to test the reliability of endogenous genes. Therefore, this study evaluated 10 reference candidate genes in two different experimental conditions in order to obtain stable genes to be used as reference in expression studies related to scrotal hernias in pigs. Two independent experiments were performed: one with 30 days-old MS115 pigs and the other with 60 days-old Landrace pigs. The inguinal ring/canal was collected, frozen and further submitted to real-time PCR analysis (qPCR). For the reference genes stability evaluation, four tools were used: GeNorm in the SLqPCR, BestKeeper, NormFinder and Comparative CT. A general ranking was generated using the BruteAggreg function of R environment. In this study, the RPL19 was one of the most reliable endogenous genes for both experiments. The breed/age effects influenced the expression stability of candidate reference genes evaluated in the inguinal ring of pigs. Therefore, this study reinforces the importance of evaluating the stability of several endogenous genes previous their use, since a consensual set of reference genes is not easily obtained. Here, two sets of genes are recommended: RPL19, RPL32 and H3F3A for 30-days MS115 and PPIA and RPL19 for the 60 days-old Landrace pigs. This is the first study using the inguinal ring tissue and the results can be useful as an indicative for other studies working with gene expression in this tissue.