The pill clams Pisidium chilense (d'Orbigny, 1846) and Pisidium huillichum (Ituarte, 1999) showed a 2C DNA content of 4.04 ± 0.4 pg and 2.35 ± 0.24 pg, respectively. The somatic DNA content of these 2 Chilean Pisidium Pfeiffer 1821 species is within the range described previously for diploid Heterodonta, despite the fact that both showed a high chromosome number (> 100–120 chromosomes) equivalent to the polyploid level described for other species of the genus. The nuclear DNA content is shown here for the first time for species of the family Sphaeriidae. The Pisidium, like other species, are not economically important as edible species, but they play a significant role in the dynamics of nutrient and energy cycles in freshwater bodies similar to that described for other Sphaeriidae species.