RESUMEN
Copepods have been widely used to evaluate toxicity of metals present in marine environments. However, a technical difficulty is to understand the possible routes of metal uptake and to identify in which tissues or organs metals are being accumulated. Traditional techniques are hard to be employed once each organ has to be analyzed separately. Autoradiography is an alternative technique to circumvent this limitation, since metal distribution in tissues can be visualized and quantified, even in small organisms like copepods. In the present study, accumulation and distribution of (64)Cu in the copepod Calanus hyperboreus was studied using autoradiography. Copepods were exposed for 2 h to copper (2.3 mg L(-1); 1.08 MBq (64)Cu mg(-1) Cu) and then allowed to depurate for 2 h in clean seawater. Total (64)Cu was determined by gamma-spectrometry after a metal exposure and a depuration period. (64)Cu distribution was determined based on images generated by autoradiography. Metal accumulation was observed on all external surfaces of the copepods, being accumulated mostly on the ventral region, followed by dorsal, urossoma and internal regions. After depuration, radioactivity levels had a decrease in the sum of external body surface. Our results show that copper uptake by C. hyperboreus is fast and that a non-negligible proportion of the accumulated metal can reach internal tissues, which may lead to detrimental physiological effects. Moreover, whole-body autoradiography was demonstrated to be an efficient technique to study copper accumulation and body distribution in a very small organism such as the copepod C. hyperboreus.
Asunto(s)
Autorradiografía/métodos , Cobre/análisis , Crustáceos/metabolismo , Contaminantes Químicos del Agua/análisis , Imagen de Cuerpo Entero/métodos , Animales , Cobre/metabolismo , Radioisótopos de Cobre/análisis , Radioisótopos de Cobre/metabolismo , Crustáceos/química , Distribución Tisular , Contaminantes Químicos del Agua/metabolismoRESUMEN
Previously, we demonstrated a higher silver body burden when Daphnia magna were exposed to silver in the presence of environmentally relevant concentrations (25 nM) of reactive sulfide, but the explanation was unclear. In the present study, D. magna were exposed to AgNO3 (0.93 microg Ag/L=8.6 nM as a mixture of cold Ag and (110m)Ag) in synthetic water in either the presence or absence of 25 nM sulfide as zinc sulfide clusters. After 1-h exposure, daphnids were transferred to clean water for up to 5-h depuration. At different times of Ag exposure and depuration, daphnids were randomly sampled for whole body silver burden. Also, after 1 h, daphnids were sampled for silver accumulation in "gills" (small organs on the thoracic appendages), digestive tract, and carcass. Other groups were exposed to the same silver and sulfide concentrations for 1 h and then sampled for whole-body autoradiography. Silver body burden was about two-fold higher in the presence of sulfide. A two-fold increase in silver burden in "gills" and digestive tract, but not in carcass, was also observed in the presence of sulfide. Absolute differences due to sulfide were greatest in digestive tract and explained most of the difference in whole body burden. Transfer to clean water caused a significant drop in silver concentration in whole body and all compartments to similar levels in the two groups after 5-h depuration. These results indicate that the higher silver body burden observed in the presence of sulfide is mainly due to sulfide-bound silver in the digestive tract of the daphnids. This conclusion is supported by autoradiography, which showed a high concentration of silver in the digestive tract of daphnids exposed to Ag/sulfide.