Standardization of molecular analysis techniques for DNA identification of bird species of the Psittacidae family

Abstract

Birds of the Psittacidae family belong to one of the groups with the most negative impact from wildlife trafficking, which has consequences beyond removing these species from the wild. This work aimed to standardize DNA extraction techniques from blood, feathers, and eggshells of Psittacidae to molecular identification and help track the place of origin of the seized bird. Blood and feather samples from adult of the Turquoise-fronted Parrot, Amazona aestiva, individuals (n = 5) were collected, and additionally, eggshells from nests (n = 3). We tested five nucleic acid extraction techniques.DNA concentrations and purity were evaluated by fluorimetry and spectrophotometry. As a result, the extraction by phenol provided a higher concentration of DNA from blood (20 µg/ml) and feathers (0.4 µg/ml), while for the eggshells, the acetate technique was the most efficient (33 µg/ml). Polymerase chain reaction amplification of mitochondrial cytochrome B (CytB) and 16 S genes were positive for all DNA samples. We obtained favorable conditions for DNA extraction from blood, feathers, and eggshells of the Turquoise-fronted Parrot using the five DNA extraction methods tested. These protocols can be used as a reference for other species of Psittacidae, and can be applied for biodiversity and forensic studies.