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 6.25ug/ 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.

Introduction: Amoebiasis is a parasitic disease caused by Entamoeba histolytica that may lead to death in developing countries. Few important risk factors have been identified in the development of amoebic liver abscess (ALA). There are limited reports that suggest an association between antigens of the major histocompatibility complex (MHC) particularly class II antigens and ALA development. This present work aimed at studying the possible association of HLA antigens with ALA and disease severity. Results of the study may serve as a guide for further immunological studies dealing with E. histolytica. Methods: This preliminary study involved two groups of subjects: 20 ALA patients in the experimental group and 40 healthy individuals in the control group. Cases were selected from adult Malay patients confirmed with ALA based on clinical signs and symptoms, radiological findings, microbiological findings and who were admitted to the medical or surgical ward, Hospital USM, Kelantan. Venous blood was obtained from each patient and HLA typing was then conducted using polymerase chain reaction specific primer sequence. Results: HLA DR12 was most frequently found in the healthy control and ALA groups at 40% and 55% respectively. HLA DQ7 and DQ8 were found to have the highest percentage in the ALA group at 65%. In the control group, HLA DQ8 (57.5%) had the highest percentage. Conclusion: HLA antigens play a role in acquisition of ALA and provide understanding of the disease outcome.