Toxicidade de cádmio e zinco em Danio rerio: comparação entre valores permitidos em legislação para proteção da vida aquática e a potencial atuação como interferentes endócrinos
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Data
2016-02-19
Autores
Alkimin, Gilberto Dias de [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A contaminação dos ambientes aquáticos por metais é uma preocupação mundial, pois são elementos bioacumulativos e persistentes. Diversos desses compostos possuem a capacidade de alterar o sistema endócrino (denominados interferentes endócrinos – IE), sendo o cádmio (Cd) e o zinco (Zn) suspeitos dessa atuação, segundo a literatura. Para verificação dessa potencialidade algumas análises biológicas são indicadas como: 1) a realização do teste de dessorção no auxílio ao posterior cálculo do fator de bioconcentração (FBC), para detecção do real valor incorporado do contaminante pelo organismo e predição dos efeitos tóxicos causados; 2) análises histológicas de órgãos como as gônadas e o fígado e 3) análise de vitelogenina (VTG) no fígado de machos. Dessa forma o objetivo deste trabalho foi averiguar a atuação endócrina dos metais Cd e Zn em concentrações permitidas em ambientes aquáticos para a proteção da vida aquática por meio de: a) estudo do tempo de dessorção de Cd e Zn da parte externa do organismo; b) determinação do FBC de Cd e Zn em D. rerio após exposição crônica; c) avaliação da possibilidade de Cd e Zn provocarem alterações histológicas em gônada e fígado de machos e fêmeas de D. rerio e d) análise da capacidade de Cd e Zn em induzir a produção de VTG no fígado de machos de D. rerio. Para a realização do teste de dessorção, organismos adultos de D. rerio, foram expostos por 72 horas a Cd (1 µg de Cd/L) e Zn (180 µg de Zn/L) seguindo-se a lavagem do organismo-teste em ácido nítrico 2% por diferentes tempos (0h; 5; 60; 90; 120; 150 e 180 minutos) e posterior construção da curva de dessorção. As análises de FBC, histologia e histoquímica foram realizadas após exposição crônica (21 dias) em regime semi-estático, com troca total da solução a cada 72 horas, em separado a concentrações permitidas por legislação nacional (1 µg de Cd/L e 180 µg de Zn/L) e internacional (0,25 µg de Cd/L e 120 µg de Zn/L) para proteção da vida aquática. Desse modo, após o final da exposição organismos foram levados a digestão, leitura da concentração do contaminante e o FBC foi calculado, bem como machos e fêmeas foram sacrificados e passaram por procedimento de preparo histológico, corados com hematoxilina e eosina e análise qualitativa de gônada e fígado e machos após preparo histológico sofreram marcação histoquímica para detecção de VTG no fígado. Pelos resultados obtidos, verificou-se que a dessorção dos metais apresentou diferença, sendo a dessorção de Cd realizada em 90 minutos e a de Zn em 60 minutos; a bioconcentração de Cd foi semelhante nas duas concentrações de exposição e a de Zn foi inversamente proporcional à concentração exposta. As análises histológicas mostraram que Cd e Zn provocam alterações de forma pronunciada tanto em gônadas quanto em fígados de fêmeas, enquanto gônadas de machos não sofreram graves alterações e os fígados apresentaram lesões que podem comprometer o funcionamento do órgão, além disso, observou-se a produção de VTG no fígado de machos de D. rerio expostos a Cd e Zn. Por fim, a união das técnicas utilizadas comprova que Cd e Zn, mesmo em concentrações permitidas por legislação são capazes de atuarem como IE em D. rerio, necessitando de mudanças na legislação para valores mais restritivos com a finalidade de garantir a saúde endócrina dos organismos aquáticos.
The contamination of aquatic environments by metals is a global concern because they are bioaccumulative and persistent elements. Several such compounds have the ability to change the endocrine system (called endocrine disruptors - ED), and cadmium (Cd) and zinc (Zn) that suspicious activity, according to the literature. To verify the potential some biological analyzes are given as: 1) the realization of the desorption test to aid the subsequent calculation of bioconcentration factor (BCF), for determining the real corporate value of the contaminant by the body and prediction of toxic effects; 2) histological analysis of organs such as the gonads and liver and 3) analysis of vitellogenin (VTG) in the liver of males. Thus the aim of this study was to determine the endocrine actions of Cd and Zn in allowed concentrations in aquatic environments for the protection of aquatic life through: a) study time desorption of Cd and Zn from outside the body; b) determination of CBF of Cd and Zn in D. rerio after chronic exposure; c) assessment of the possibility of Cd and Zn cause histological changes in the liver and gonads of males and females of D. rerio d) analysis of the ability of Cd and Zn in inducing the production of VTG in the liver of males of D. rerio. To perform the desorption test, adult organisms of D. rerio were exposed for 72 hours Cd (1 µg/L of Cd) and zinc (180 µg/L of Zn) followed by washing of the organism under test 2% nitric acid for different times (0 h, 5, 60, 90, 120, 150 and 180 minutes) and subsequent construction of desorption curve. The analyses of BCF, histologic and histochemical were performed after chronic exposure (21 days) under semi-static system, with complete exchange of the solution every 72 hours in separat concentrations allowed by national legislation (1 µg of Cd/L and 180 µg of Zn/L) and international (0.25 µg Cd/L and 120 µg Zn/L) for protection of aquatic life. Thus, after the end of exposure organisms were taken digestion, of the contaminant concentration of the reading and the CBF was calculated as well as males and females were euthanized and underwent histological preparation procedure, stained with hematoxylin and eosin and qualitative analysis of gonads and liver histological was performed and males after preparing suffered histochemical staining VTG detection in the liver. This waythe desorption of metals it was different, with desorption of Cd performed in 90 minutes and Zn in 60 minutes, the bioconcentration of Cd was similar in both exposure concentration and the Zn was inversely proportional to the concentration exposed, histological analyzes show that Cd and Zn cause pronounced changes in both gonads, as in female livers, but the gonads of male suffered no serious changes and livers showed lesions that can compromise the function of organ, besides, it was observed VTG production in the liver D. rerio males exposed to Cd and Zn. Finally, the union of techniques shows that Cd and Zn, even at concentrations allowed by law are able to act as IE in D. rerio, requiring changes in legislation for more stringent values in order to ensure the endocrine health organisms water.
The contamination of aquatic environments by metals is a global concern because they are bioaccumulative and persistent elements. Several such compounds have the ability to change the endocrine system (called endocrine disruptors - ED), and cadmium (Cd) and zinc (Zn) that suspicious activity, according to the literature. To verify the potential some biological analyzes are given as: 1) the realization of the desorption test to aid the subsequent calculation of bioconcentration factor (BCF), for determining the real corporate value of the contaminant by the body and prediction of toxic effects; 2) histological analysis of organs such as the gonads and liver and 3) analysis of vitellogenin (VTG) in the liver of males. Thus the aim of this study was to determine the endocrine actions of Cd and Zn in allowed concentrations in aquatic environments for the protection of aquatic life through: a) study time desorption of Cd and Zn from outside the body; b) determination of CBF of Cd and Zn in D. rerio after chronic exposure; c) assessment of the possibility of Cd and Zn cause histological changes in the liver and gonads of males and females of D. rerio d) analysis of the ability of Cd and Zn in inducing the production of VTG in the liver of males of D. rerio. To perform the desorption test, adult organisms of D. rerio were exposed for 72 hours Cd (1 µg/L of Cd) and zinc (180 µg/L of Zn) followed by washing of the organism under test 2% nitric acid for different times (0 h, 5, 60, 90, 120, 150 and 180 minutes) and subsequent construction of desorption curve. The analyses of BCF, histologic and histochemical were performed after chronic exposure (21 days) under semi-static system, with complete exchange of the solution every 72 hours in separat concentrations allowed by national legislation (1 µg of Cd/L and 180 µg of Zn/L) and international (0.25 µg Cd/L and 120 µg Zn/L) for protection of aquatic life. Thus, after the end of exposure organisms were taken digestion, of the contaminant concentration of the reading and the CBF was calculated as well as males and females were euthanized and underwent histological preparation procedure, stained with hematoxylin and eosin and qualitative analysis of gonads and liver histological was performed and males after preparing suffered histochemical staining VTG detection in the liver. This waythe desorption of metals it was different, with desorption of Cd performed in 90 minutes and Zn in 60 minutes, the bioconcentration of Cd was similar in both exposure concentration and the Zn was inversely proportional to the concentration exposed, histological analyzes show that Cd and Zn cause pronounced changes in both gonads, as in female livers, but the gonads of male suffered no serious changes and livers showed lesions that can compromise the function of organ, besides, it was observed VTG production in the liver D. rerio males exposed to Cd and Zn. Finally, the union of techniques shows that Cd and Zn, even at concentrations allowed by law are able to act as IE in D. rerio, requiring changes in legislation for more stringent values in order to ensure the endocrine health organisms water.
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Palavras-chave
Metais potencialmente tóxicos, Dessorção, Fator de bioconcentração, Histologia, Histoquímica, Potentially toxic metals, Desorption, Bioconcentration factor, Histology, Histochemistry