Two species of Callithrix, C. jacchus (Linnaeus, 1758) and C. penicillata (É. Geoffroy, 1812), are considered invasive in Rio de Janeiro. This study determined the genetic and morphological diversity and verified the species involved in the hybridization of 10 individuals from the municipalities of Silva Jardim (N = 9) and Rio das Ostras (N = 1). We compared the external morphology and skull of C. jacchus (N = 15) and C. penicillata (N = 14) specimens deposited in the collection of the National Museum of Rio de Janeiro (MN- UFRJ). Phylogenetic (maximum likelihood and Bayesian inference) and phylogeographical analyses (network analysis) were performed based on cytochrome b sequences. These analyses included hybrids from the metropolitan region of Rio de Janeiro (N = 3), C. penicillata (N = 2), C. jacchus (N = 2), C. geoffroyi (N = 2), C. kuhlii (N = 2), C. aurita (N = 1), and as outgroups, Mico emiliae (N = 1) and Saguinus mystax (N = 1). The pelage and skull characters of most hybrids were more closely related to C. jacchus. Skull morphometric analysis revealed an intermediate state for the hybrids. Phylogenetic analyses revealed a high similarity between the hybrids and C. penicillata. Six haplotypes of hybrids were identified. Network analysis including them and C. penicillata recovered the topology generated by phylogenetic analysis. The results corroborate that C. jacchus and C. penicillata participate in the hybridization process. There was no geographic structure between hybrids from the coastal lowlands and from the metropolitan region of Rio de Janeiro.