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A modified diphasic liver infusion agar medium was prepared manually in the laboratory in order to be used for culturing Entamoeba histolytica in vitro. This medium is a good alternative for the commercial one and has the main ingredients for the parasite's growth. E. histolytica was successfully cultured in the present study and PCR technique was performed to confirm Entamoeba species. The growth of E. histolytica trophozoites had started in the third day post-cultivation and reached its highest rate in the seventh. Then the growth started to decrease until the trophozoites completely disappeared after 12-14 days post-cultivation. Cryopreservation using glycerol 10% (V/V) was used in order to preserve trophozoites for later cultivation.
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