The mixture of Green Malachite (MG) and formalin is the most effective chemical used to treat Ichthyophthirius multifiliis and other prasites infecting fish.However, MG was prohibited to usein aquaculture due to its potential risks to the environment, animal and human health. The prohibited use of MG has caused many difficulties to treat this parasite and other aquatic diseases. This study was conducted to examine a new, environmental friendly, safe and effectively method alternative to MG. Formalin 25 ppm (F25) was combined with a high oxidizing PHA (15% peracetic acid, 25% hydrogen peroxideand 25% acetic acid) at different concentrations of 1, 3, 5, 7, 10 ppm (PHA1, PHA3, PHA5, PHA7, PHA10) and the mixture was used to experimentally treat theronts (in vitro), and mature Ichthyophthirius multifiliis (trophonts) parasitized on Grass carp (in vivo), following with comparison to mixture of Green malachite and formalin (F25 + G0.25). The in vitrotest results show that the mixtureof F25 + PHA10 could kill 96.5% theronts within 48h. For in vivotest, treatments were conducted 72h post infection. After 10 days of experimental treatment, the control group (infected without treatment) had a high mortality rate (85.56%).In contrast, the F25 + G0.25 showed high efficiency in controlling this parasite with 94.43% fish survival. The mixtures F + PHA were effective to mitigate mortality rate compared to the control group, in which the mixture of F25 + PHA10 resulted in highest survival rate (92.23%), following with low prevalence and infection intensity, 5.13 and 1.81 parasites/sample at 40 x magnification, respectively. In addition, Ichthyophthirius multifiliis exhibited changes in morphology and moving behaviour. Results of following 10 days post treatment showed that fish treated with F25 + PHA10 recoveredeffectively, stopped mortality and probably created immunity against this parasite.