Colour patterns are diverse in trichomycterine catfishes and are often used to diagnose species. Here, we analyse the first case of adults of two syntopic species of Trichomycterus sharing nearly identical colour patterns: a rare new species of the subgenus Paracambeva and Trichomycterus maculosus, a distantly related species of the subgenus Trichomycterus. Both species are endemic to the upper Rio Paraíba do Sul basin (RPSB), which had a different course until the Tertiary period and is situated within the Southeastern Brazilian Continental Rift, mostly active in the Eocene-Oligocene. A time-calibrated multigene analysis, 3144 bp, supported the new species as sister to Trichomycterus itatiayae, both comprising a lineage with Middle Miocene age, when that colour pattern would have first arisen. The new species is diagnosed by characters from the latero-sensory system and bone morphology. Our results, combined with available biogeographical data, indicated the colour pattern of T. maculosus arising in the Late Pliocene, following the dispersal of its group to the upper RPSB after river course changing. Two hypotheses for the independent origin of the same colour pattern are discussed. First, a case of evolutionary convergence for adaptation to live on a similarly coloured gravel substrate, giving some cryptic advantage against predators. Second, mimetic association through anti-predation features. In the latter case, although trichomycterids lack fin spines to inoculate venom as in other catfishes, the species here studied have a supposed axillary gland above the pectoral fin, just posterior to the opercular odontodes, but with properties and functions still unknown.
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