Black rats (Rattus rattus) are native to the Indian subcontinent but have now colonized most continents and islands following human movements and international trade. They are involved in the circulation and transmission to humans of many zoonotic agents as well as in massive damage to food stocks and native biodiversity in the regions they have settled. This study investigates the genetic diversity and possible origins of black rats from Benin, West Africa. We sequenced the complete mitochondrial cytochrome b gene in 90 individuals from nine localities in Benin. These sequences were subsequently compared to 390 other cytochrome b haplotypes from individuals from various European, Asian, American and African localities. Nucleotide polymorphism analysis, haplotype network and maximum likelihood phylogenetic tree reconstructions showed low mitochondrial diversity in black rats from Benin. Our results also suggest at least two distinct introduction events: one introduction probably occurred during the spice trade (15th-17th century) through the Indies Road connecting Europe to Asia. Other introduction events could have occurred more recently following the intensification of globalized trade from the eighteenth century, and onwards.