Blastocystis infection accounts for one of the causes of gastrointestinal problems with the prevalence rate of 3-100% worldwide. There is a wide range of drugs examined for the treatment of infected patients, among them metronidazole (MTZ) has been introduced as one of the efficient drugs. Besides to the suitable clinical effects, the administration of MTZ has some reported side-effects which emphasize on the identification of putative alternates. To this end, we aimed to evaluate the cytotoxicity effect of a newly-introduced synthetic antimicrobial peptide (AMP) named CM11 on in vitro cultured Blastocystis. Our results exhibited that CM11 treatment affected the viability of parasites in two cultural conditions including culturing alone and in co-culture with the Caco-2 cell line. The time- and dose-dependent effect of CM11 was consistent with the effect of MTZ which was used as control positive. The highest toxicity effect of CM11 was observed at the concentration of 24 μg/ml, leading to 28.7% and 25% viable parasites after 24 h and 48 h incubation times, respectively. Interestingly, the disruption of the Blastocystis cell membrane could be observed in the treated parasites. Therefore, CM11 can be suggested as a potential treatment for Blastocystis-infected patients after further in vitro and in vivo assessments.