Ixodes amaraliFonseca, 1935 is a tick species endemic to Brazil, where it has been reported only in the Caatinga (semiarid), Cerrado (savannah), and Atlantic forest biomes. While females of I. amarali have been several times collected, the male has remained undescribed. Here we report new collections of larva, nymph, females, and one male of I. amarali from the Pará State, northern Brazil, which constitute the first records of this tick species in the Amazon biome. The male of I. amarali is morphologically described. Partial sequences of the mitochondrial 16S rRNA gene were generated from the single male and one female of I. amarali from the Amazon biome. Phylogenetic analysis inferred from 16S rRNA partial sequences showed that I. amarali (male and female) from Amazon grouped with a sequence of I. amarali from southeastern Brazil. These three sequences clustered into a monophyletic group with sequences of Ixodes loricatus Neumann, 1899, Ixodes luciae Sénevet, 1940, and Ixodes schulzei Aragão and Fonseca, 1951, which constitute the so-called “I. loricatus group”. Morphological characters to separate the male of I. amarali from I. loricatus, and I. luciae are provided.