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Currently, the double staining method for bone and cartilaginous staining using alizarin red and cyan blue, respectively, generates high costs in the implementation of protocols for anatomy and systematic studies in vertebrates. The objective of this work was to compare and analyze the dye extracted from Bixa Orellana seeds and the alizarin red used in bone staining in Piaractus Brachypomus juvenile fish. 70 juvenile fish with 2 ± 0.4 cm total length were used; they were sedated using 2-phenoxiethanol at a final concentration of 500 mg/L and slaughtered under the standards and procedures for laboratory animals. They subsequently underwent a series of tissue conservation phases. 35 fish were allocated to each treatment (alizarin red and annatto) and bone structures stained with each dye were observed and identified. ie (intensity of emerging light) and id (integral density) were also analyzed in diaphanization specimens for each treatment. No significant differences were observed for ie (P = 0.10), in contrast to the id (p < 0.05) for the vertebral column and dorsal fin. In conclusion Bixa Orellana (annatto) has dying affinity in bone structures of juvenile fish and is therefore a good alternative for the diaphanization technique and osteological studies of vertebrates at a low cost.

Método alternativo de tinción ósea en peces juveniles empleando extracto de Bixa orellana