The mode of action of amitraz is thought to be its toxic effects on a receptor for a neuromodulator, octopamine. Resistance could arise from modifications of this receptor so that it would not be affected by amitraz. A putative octopamine receptor cDNA was cloned and sequenced from a cattle tick in Australia. However, when the sequence was compared between Australian strains of amitraz-susceptible and resistant ticks, no differences were detected. We have sequenced this putative octopamine receptor gene in tick strains from America. The American ticks have a sequence almost identical to that of the Australian ticks with no deletions or additions in the open reading frame. In a Brazilian strain and a Mexican strain that are very resistant to amitraz, there are two nucleotide substitutions that result in amino acids different from all the susceptible strains. Discovery of these mutations only in amitraz-resistant ticks provides the first evidence for the possibility of an altered pesticide target site as a mechanism of amitraz resistance in ticks.