Metal bioaccumulation and toxicity to aquatic organisms depends on factors such as magnitude, duration and frequency of the exposure. The type of the exposure affects the toxicokinetic processes in the organisms. In this study, we carried out 30-day toxicity tests on juveniles of Ruditapes philippinarum exposed to increasing, continuous and pulsed exposure. Organisms were exposed to copper-spiked sediments followed by a 10-day recovery period. We assessed the interaction between the kinetics of subcellular copper partitioning and the growth response. Results showed that the growth rate of the bivalve was inversely correlated to the bioaccumulation rate and that sublethal copper concentrations stimulated the detoxification mechanisms inside the organism regardless the type of the exposure. However, a large stimulatory effect on growth was observed during the recovery period, associated with significant negative accumulation rate values and dependent on the type of antecedent exposure. This suggested that on individual and short-term basis pulsed exposures have a more adverse effect compared to increasing or continuous exposure scenarios.
Histological techniques are a traditional and essential tool for adequately comprehending cellular morphology and physiology in several scientific fields, including ecotoxicological studies. Although bioassays generally recommend using small test organisms, based on advantages such as low maintenance costs and the possibility of a higher number of replicates, the histological process of such organisms is still challenging due to the difficulty of handling them. The present study proposes an accessible protocol for performing the histology of a biological sample of 8 mm maximum length and using histopathological biomarkers in Senegal sole larvae for ecotoxicological tests. Eight-day old larvae of Solea senegalensis obtained from breeders’ natural spawns in cultivation conditions were submitted to histological processing by an adjusted protocol comprising fixation, agarose array pre-embedding, dehydration, paraffin embedding, slides’ preparation, and image analysis by light optical microscope coupled to a digital camera. Low-cost and easy-to-use alternatives were applied in the technique execution. The histological steps were detailed, and S. senegalensis morphological features were presented. This can be considered valuable background information for ecotoxicological studies where changes in cell morphology and physiology are expected.
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