Induction of cell cytotoxicity during Trichomonas tenax incubation with S-G cells for 24 h. (A) The cell viability of S-G cells incubated with T. tenax at different MOIs (1, 2, 4, 8) for 6, 12, and 24 h was assessed using a microscope (Yellow arrow: aggregated T. tenax, white arrow: round S-G cell shape, Scale Bar: 50 μm). (B) The CPE of S-G cells co-incubated with T. tenax was observed by staining with Giemsa buffer in 24-well plated. (C) Stained cells were calculated by ImageJ. (D) Lactate dehydrogenase (LDH) cytotoxicity assay was performed on S-G cells after coincubation with T. tenax at different MOIs for 24 h (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, MOI: multiplicity of infection).