The little fire ant, Wasmannia auropunctata, is native to Central America, but has been introduced into many parts of the world. We examined the current distribution of W. auropunctata in Cameroon, tested for aggression between workers from different parts of the country, and examined the genotypes of workers, queens, and males to evaluate the mating system. We found W. auropunctata at 36 sites in three provinces (Centre, East, and South). We found W. auropunctata only in human-disturbed habitats. Its spread appears to be primarily human mediated. Aggressive behaviour was almost non-existent between workers from different sites, indicating that there is only one supercolony in Cameroon. Our genetic analysis found that only one male/female pair of clones was introduced into Cameroon, probably from Gabon. No new male clonal lineage was identified, whereas new sexually derived female clonal lineages were noted. Apart from the genotype of the founding queen, which was well distributed but generally not dominant, a new clonal queen genotype emerged and was both omnipresent and dominant at most sites. These results may be useful in the development of management strategies.