Comparison of the Anaerocult A and the oil blocking methods for the in vitro cultivation of Entamoeba histolytica

Entamoeba histolytica, the causative agent for human amoebiasis, is among the most deadly parasites, accounting for the second highest mortality rate among parasitic diseases. Because this parasite dwells in low oxygen tension, for its cultivation, microaerophilic conditions are required to mimick the human gut environment. Several methods developed for optimal growth environment are commercially available and some are conventionally modified in-house which include the Anaerocult A and oil blocking preparation methods. This study was undertaken to compare the reliability of the Anaerocult A and the oil blocking methods in generating anaerobic environment for cultivation of E. histolytica. The trophozoites of E. histolytica HM1: IMSS strains were axenically cultivated in TYI-S-33 medium in culture incubated anaerobically by using Anaerocult A (Merck) and mineral oil blocking method. The outcomes of both methods were determined by the minimum inhibitory concentration (MIC) of metronidazole against E. histolytica by giving a score to the growth pattern of the trophozoites. The reliability of both methods was assessed based on susceptibility testing of E. histolytica to metronidazole. The MIC obtained by both anaerobic condition methods was 6.25 ug/ ml, thus showing that oil-blocking method is comparable to the Anaerocult A method and therefore, considered as a reliable method for generating an anaerobic environment for the cultivation of E. histolytica.