The pygmy marmoset, the smallest of the anthropoid primates, has a broad distribution in Western Amazonia. Recent studies using molecular and morphological data have identified two distinct species separated by the Napo and Solimões-Amazonas rivers. However, reconciling this new biological evidence with current taxonomy, i.e., two subspecies,
Cebuella pygmaea pygmaea
(Spix, 1823) and
Cebuella pygmaea niveiventris
(Lönnberg, 1940), was problematic given the uncertainty as to whether Spix’s pygmy marmoset (
Cebuella pygmaea pygmaea
) was collected north or south of the Napo and Solimões-Amazonas rivers, making it unclear to which of the two newly revealed species the name
pygmaea
would apply. Here, we present the first molecular data from Spix’s type specimen of
Cebuella pygmaea
, as well as novel mitochondrial genomes from modern pygmy marmosets sampled near the type locality (Tabatinga) on both sides of the river. With these data, we can confirm the correct names of the two species identified, i.e.,
C. pygmaea
for animals north of the Napo and Solimões-Amazonas rivers and
C. niveiventris
for animals south of these two rivers. Phylogenetic analyses of the novel genetic data placed into the context of cytochrome
b
gene sequences from across the range of pygmy marmosets further led us to re-evaluate the geographical distribution for the two
Cebuella
species. We dated the split of these two species to 2.54 million years ago. We discuss additional, more recent, subdivisions within each lineage, as well as potential contact zones between the two species in the headwaters of these rivers.