Stocks of the Tetrahymena pyriformis complex have been collected in North America and their mating reactivity has been studied. In addition to stocks mating with Tetrahymena americanis, T. borealis, T. pigmentosa, T. hyperangularis, and T. australis, stocks belonging to old syngen 5 and three new mating groups, numbers 13, 14, and 15, were discovered. Syngen 5 and groups 13 and 14 are distinct “biological” species, based on their reproductive isolation from other groups and on the ability of withingroup crosses to produce immature progeny. These species have been named T. hegewischi n. sp., T. sonneborni n. sp., and T. nipissingi n. sp., respectively. The cross between the two group 15 stocks did not produce immature progeny, and there is not sufficient evidence to conclude that this pair of stocks represents a separate species. Temperature tolerance measurements have been made on stocks representing all known micronucleate members of “pyriformis” complex. Within each species, the range of temperature tolerances is narrow; the average within‐species standard deviation is 0.63°C. The species averages range from 32.7 to 40.7°C. Using syngen numbers, the order from lowest to highest temperature tolerance is 9, 8, 10, 7, 6, 4, 13, 14, 12, 11, 5, 3, 2, 1. The large differences among species make temperature tolerance a useful aid in identification, but the origins of the differences among species are unknown.