Termite colonies are usually headed by primary reproductives,which establish nests during the swarming season.However,secondary reproductives may develop in some species and become supplementary or replacement breeders,extending colony lifespan.Here we investigate the developmental origin,fecundity and genetic characterization of ergatoid reproductives in the Neotropical termite Nasutitermes aquilinus (Holmgren),using morphometrical and histological techniques,five microsatellite loci and the COI mitochondrial DNA.Twelve measurements performed on 208 apterous individuals of N.aquilinus revealed 10 groups,including ergatoid females,which developed from major workers through two successive molts,and were characterized by the presence ofimaginal features such as eyes and wing buds.The differentiation of these features was correlated to physogastric development in these ergatoids.Histology revealed oocytes in all maturation stages in worker-derived reproductives of N.aquilinus,presence of nonflagellate spermatozoa inside the spermatheca,and royal fat body.Thus,ergatoid reproductives were reproductively functional.According to the genotypes of 221 individuals from 11 nests,and mitochondrial haplotypes of 43 ergatoids,73％ of the colonies were simple families,whereas 27％ were extended families.Despite the occurrence of related reproductives,low inbreeding rates were detected within and among colonies.Such values could be explained given that sib mating itself cannot result in a higher inbreeding rate but depend on several factors discussed in detail.This is the first study to investigate the genetic structure of termite colonies influenced by the development ofergatoids,and further investigations are encouraged to understand the influence of these reproductives on colony lifespan.