Blepharismins are photodynamic hypericin‐like dianthrones produced as a variable pigment blend in Blepharisma ciliates and mostly studied in the Afro‐Asiatic Blepharisma japonicum. The present work describes the bioactivity of pigments from the Brazilian Blepharisma sinuosum. Comparative analyses showed that the pigments from both species can trigger photo‐induced modifications in phospholipids, but different redox properties and biological activities were assigned for each pigment blend. Stronger activities were detected for B. sinuosum pigments, with the lethal concentration LC50 10 × lower than B. japonicum pigments in light‐irradiated tests against Bacillus cereus and less than half for treatments on the human HeLa tumor cells. HPLC showed B. sinuosum producing a simpler pigment blend, mostly with the blepharismin‐C (~ 70%) and blepharismin‐E (~ 30%) types. Each blepharismin engaged a specific dose–response profile on sensitive cells. The blepharismin‐B and blepharismin‐C were the most toxic pigments, showing LC50 ~ 2.5–3.0 µm and ~ 100 µm on B. cereus and HeLa cells, respectively, after illumination. Similarity clustering analysis compiling the bioactivity data revealed two groups of blepharismins: the most active, B and C, and the less active, A, D and E. The B. sinuosum pigment blend includes one representative of each clade. Functional and medical implications are discussed.