Invertebrate infection models that can be evaluated at human body temperature are limited. In this study, we utilized the two-spotted cricket, a heat-tolerant insect, as an animal infection model of human pathogenic fungi. Injection of human pathogenic fungi, including Candida albicans, Candida glabrata, and Cryptococcus neoformans killed crickets within 48 h at both 27˚C and 37˚C. The median lethal dose values (LD50 values) of C. albicans and C. glabrata against crickets were decreased at 37˚C compared to that at 27˚C, whereas the LD50 value of C. neoformans was not different between 27˚C and 37˚C. Heat-killed cells of the three different fungi also killed crickets, but the LD50 value of the heat-killed cells was higher than 5-fold that of live fungal cells in the respective species. C. neoformans gene-knockout strains of cna1, gpa1, and pka1, which are required for virulence in mammals, had greater LD50 values than the parent strain in crickets. These findings suggest that the two-spotted cricket is a valuable infection model of human pathogenic fungi that can be used to evaluate fungal virulence at variable temperatures, including 37˚C, and that the killing abilities of C. albicans and C. glabrata against animals are increased at 37˚C.