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Due to morphological similarity among females, molecular tools are essential. Although\n                      <jats:italic>cytochrome c oxidase subunit 1<\/jats:italic>\n                      (\n                      <jats:italic>COI<\/jats:italic>\n                      ) is the standard animal barcode, its limited resolution in recently diverged species necessitates more informative regions, such as\n                      <jats:italic>cytochrome b<\/jats:italic>\n                      (\n                      <jats:italic>CYTB<\/jats:italic>\n                      ) and\n                      <jats:italic>NADH dehydrogenase 2<\/jats:italic>\n                      (\n                      <jats:italic>ND2<\/jats:italic>\n                      ), which are effective for Thraupidae. The aim of this study was to provide molecular mitochondrial markers to support species identification and development of tools that contribute to conservation of\n                      <jats:italic>Sporophila<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods and results<\/jats:title>\n                    <jats:p>\n                      The markers were validated using 62 adult males (13 species) and applied to 47 unidentified females from the Centro de Triagem de Animais Silvestres (CETAS). DNA was amplified via PCR, targeting\n                      <jats:italic>CYTB<\/jats:italic>\n                      and\n                      <jats:italic>ND2<\/jats:italic>\n                      , using genus-specific primers for all samples.\n                      <jats:italic>COI<\/jats:italic>\n                      was amplified exclusively for males, using primers based on literature. Sequences were analyzed using an ABI3500.Maximum likelihood trees and haplotype networks compared each studied region. Nearest-neighbor and barcode gap analyses confirmed marker efficiency for species identification, with\n                      <jats:italic>CYTB<\/jats:italic>\n                      and\n                      <jats:italic>COI<\/jats:italic>\n                      demonstrating the highest reliability. Among the 13 species evaluated, five showed barcode gaps across all markers, while two depended on the marker used. Female samples achieved a 100% amplification rate, with 74.47% showing high identity (&gt;\u200998%); conclusively, 52.13% were identified with high confidence, whereas the remainder belonged to species falling outside our markers\u2019 current discrimination efficiency.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      This study provides efficient tools for future research and conservation initiatives targeting\n                      <jats:italic>Sporophila<\/jats:italic>\n                      species.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s11033-026-12530-2","type":"journal-article","created":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T09:40:56Z","timestamp":1786009256000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["DNA barcode markers for birds of the genus Sporophila: advances in species identification"],"prefix":"10.1007","volume":"53","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-5781-0106","authenticated-orcid":false,"given":"J\u00fallia Costa","family":"dos Reis","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6873-7322","authenticated-orcid":false,"given":"Amanda Alves","family":"de Melo-Ximenes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7770-7707","authenticated-orcid":false,"given":"Rhewter","family":"Nunes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-1082-3632","authenticated-orcid":false,"given":"Luiz Alfredo Martins","family":"Baptista","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0390-6065","authenticated-orcid":false,"given":"Leo Caetano Fernandes","family":"Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9023-0007","authenticated-orcid":false,"given":"Mariana Pires","family":"de Campos Telles","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,6]]},"reference":[{"key":"12530_CR1","doi-asserted-by":"publisher","first-page":"653","DOI":"10.1007\/s10531-016-1263-y","volume":"26","author":"J Li","year":"2017","unstructured":"Li J, Cui Y, Jiang J, Yu J et al (2017) Applying DNA barcoding to conservation practice: a case study of endangered birds and large mammals in China. 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Every sampling procedure was conducted under the supervision of a licensed veterinarian. We adhered to the standardized methodologies for avian sample acquisition and management established by Unigen, a nationally recognized laboratory specializing in veterinary genetic diagnostics. The instructions followed are available on their website: (\n                      \n                      ) on the tab \u201cComo coletar sangue ou penas para sexagem\u201d.","order":2,"name":"Ethics","label":"Ethics approval","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"1347"}}