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Typhlodromus phialatus and T. exhilaratus are morphologically close species. Their differentiation is based on the shape of the insemination apparatus and on idiosomal setae length. However, the setae length values are often intermediate between these two species and do not allow accurate identification. Furthermore, the handful of differences in insemination apparatus shape are also questionable as a means of differentiation. Synonymy between these species has thus been questioned. Three experiments were carried out. Idiosomal seta length measurements, molecular tests and cross experiments were conducted for three populations, identified as T. exhilaratus and T. phialatus according to the shape of their insemination apparatus. The results show that the variation range of seta lengths is great and that these criteria do not allow accurate separation of these populations into two species. However, molecular tests show a species-level nucleotide differentiation between them. Cross experiments confirm this result, showing complete reproduction incompatibility between the mites bearing different insemination apparatus shapes. Therefore, T. exhilaratus and T. phialatus could be considered to be two valid species and the insemination apparatus could be considered as a pertinent diagnostic criterion at the specific level.