Ticks (Ixodida) are ecologically important ectoparasites that may impact human health and economic activities. Parasitoid wasps are natural enemies of ticks and a potential option for its biological control. In the present study, we recorded parasitism of the nymphs of the tick Amblyomma nodosum by the parasitoid wasp Ixodiphagus hookeri. The ticks, in turn, were parasitizing white-bearded manakins, Manacus manacus (Passeriformes: Pipridae) in the União Biological Reserve, one of the last major remnants of lowland rainforest in Rio de Janeiro state, southeastern Brazil. During the collection of ectoparasites from two manakins in this reserve, 20-30 parasitoid wasps hatched spontaneously from two of the ticks collected. The species of the tick nymphs and adult wasps were identified using morphological traits and molecular analyses. This record is the first documented case of I. hookeri parasitizing A. nodosum and the first time that parasitoid wasps have been found in ticks parasitizing birds in Brazil. This is also the first record of Ixodiphagus from southeastern Brazil in more than a century, with the only other record dating back to 1914. The record presented here provides valuable new information on the biology of this tick and its parasitoids, and new insights into this interspecific interaction. Further research on these vertebrate-tick-wasp interactions will be necessary to better understand the respective roles of the organisms involved in these processes.