Wolbachia pipientis is a maternally transmitted bacterium that often alters the life history of its insect host to maximize transmission to subsequent generations. Here we report on the frequency and distribution of Wolbachia infection in a widespread invasive species, the Argentine ant (Linepithema humile). We screened 1175 individual Argentine ants from 89 nests on five continents and several islands, including numerous locations in both the native (South American) and introduced ranges. We detected Wolbachia in four of 11 native populations, but only one of 21 introduced populations was infected. In the Argentine ant's native range, the distribution of Wolbachia supergroups A and B was nonoverlapping. By coupling infection frequency data with behaviourally defined colony boundaries, we show that infected and uninfected colonies are often adjacent to one another, supporting the proposition that little female-mediated gene flow occurs among Argentine ant colonies. We also conduct a phylogenetic analysis, and show that the Wolbachia infecting both native and introduced populations of Argentine ants belong to two lineages that appear to be specialized on infecting New World ants. One other lineage of Wolbachia has undergone frequent, recent episodes of horizontal transmission between distantly related, introduced insect hosts.