Evaluating the factors that promote invasive ant abundance is critical to assess their ecological impact and inform their management. Many invasive ant species show reduced nestmate recognition and an absence of boundaries between unrelated nests, which allow populations to achieve greater densities due to reduced intraspecific competition. We examined nestmate discrimination and colony boundaries in introduced populations of the red imported fire ant (Solenopsis invicta; hereafter, fire ant). Fire ants occur in two social forms: monogyne (colonies with a single egg-laying queen) and polygyne (colonies with multiple egg-laying queens). In contrast with monogyne nests, polygyne nests are thought to be interconnected due to the reduced antagonism between non-nestmate polygyne workers, perhaps because polygyne workers habituate the colony to an odor unique to Gp-9b-carrying adults. However, colony boundaries and nestmate discrimination are poorly documented, particularly for worker-brood interactions. To delimit boundaries between field colonies, we correlated the exchange of a 15N-glycine tracer dissolved in a sucrose solution with social form. We also evaluated nestmate discrimination between polygyne workers and larvae in the laboratory. Counter to our expectations, polygyne colonies behaved identically to monogyne colonies, suggesting both social forms maintain strict colony boundaries. Polygyne workers also preferentially fed larval nestmates and may have selectively cannibalized non-nestmates. The levels of relatedness among workers in polygyne colonies was higher than those previously reported in North America (mean ±SE: 0.269 ± 0.037). Our study highlights the importance of combining genetic analyses with direct quantification of resource exchange to better understand the factors influencing ant invasions.

Unicoloniality, or the absence of behavioral boundaries between nests, is thought to promote ant abundance due to reduced intraspecific competition. Workers within unicolonial populations may increase their own inclusive fitness by preferentially caring for more related individuals (nepotism), but nepotism has only rarely been documented in ants. We tested for unicoloniality and nepotism in polygyne red imported fire ants (Solenopsis invicta; hereafter fire ants). Fire ants occur in two social forms: monogyne (i.e., colonies with a single egg-laying queen) and polygyne (i.e., colonies with multiple egg-laying queens). Introduced populations of polygyne fire ants are commonly referred to as unicolonial, but cooperation between and within colonies is poorly documented. To delimit boundaries between colonies in the field, we quantified the exchange of a 15N-glycine tracer dissolved in a sucrose solution and correlated this exchange with colony genetic structure. We also quantified within-colony conflict between workers and larvae using close siblings (i.e., from the same mother) and non-siblings (i.e., from a different mother). Counter to our expectations, polygyne colonies did not exchange resources or workers, indicating distinct colony boundaries. Polygyne workers also preferentially fed larval sibling and may have preferentially cannibalized non-siblings. Polygyne colony behavior was correlated with higher levels of within-mound relatedness between workers in the field than those previously reported in North America (mean ± SE: 0.269 ± 0.037). Our study challenges fundamental assumptions about introduced populations of polygyne fire ants and suggests that polygyne colonies are multicolonial and likely engage in high levels of intraspecific competition.