Prevalence and Risk Factors Associated with Entamoeba histolytica/dispar/moshkovskii Infection among Three Orang Asli Ethnic Groups in Malaysia Tengku Shahrul Anuar 1 , Hesham M. Al-Mekhlafi 2 , Mohamed Kamel Abdul Ghani 3 , Emelia Osman 1 , Azlin Mohd Yasin 1 , Anisah Nordin 1 , Siti Nor Azreen 1 , Fatmah Md Salleh 1 , Nuraffini Ghazali 1 , Mekadina Bernadus 1 , Norhayati Moktar 1 * 1 Department of Parasitology and Medical Entomology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia, 2 Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia, 3 Programme of Biomedical Sciences, School of Diagnostic and Applied Health Sciences, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia Abstract Background: Entamoeba histolytica/Entamoeba dispar/Entamoeba moshkovskii infection is still prevalent in rural Malaysia especially among Orang Asli communities. Currently, information on prevalence of this infection among different ethnic groups of Orang Asli is unavailable in Malaysia. To contribute to a better comprehension of the epidemiology of this infection, a cross-sectional study aimed at providing the first documented data on the prevalence and risk factors associated with E. histolytica/E. dispar/E. moshkovskii infection was carried out among three Orang Asli ethnic groups (Proto-Malay, Negrito, and Senoi) in selected villages in Negeri Sembilan, Perak, and Pahang states, Malaysia. Methods/Findings: Faecal samples were examined by formalin-ether sedimentation and trichrome staining techniques. Of 500 individuals, 8.7% (13/150) of Proto-Malay, 29.5% (41/139) of Negrito, and 18.5% (39/211) of Senoi were positive for E. histolytica/E. dispar/E. moshkovskii, respectively. The prevalence of this infection showed an age-dependency relationship, with higher rates observed among those aged less than 15 years in all ethnic groups studied. Multivariate analysis confirmed that not washing hands after playing with soils or gardening and presence of other family members infected with E. histolytica/E. dispar/E. moshkovskii were significant risk factors of infection among all ethnic groups. However, eating with hands, the consumption of raw vegetables, and close contact with domestic animals were identified as significant risk factors in Senoi. Conclusions: Essentially, the findings highlighted that E. histolytica/E. dispar/E. moshkovskii parasites are still prevalent in Malaysia. Further studies using molecular approaches to distinguish the morphologically identical species of pathogenic, E. histolytica from the non-pathogenic, E. dispar and E. moshkovskii are needed. The establishment of such data will be beneficial for the public health authorities in the planning and implementation of specific prevention and control strategies of this infection in different Orang Asli ethnic groups in Malaysia. Citation: Shahrul Anuar T, Al-Mekhlafi HM, Abdul Ghani MK, Osman E, Mohd Yasin A, et al. (2012) Prevalence and Risk Factors Associated with Entamoeba histolytica/dispar/moshkovskii Infection among Three Orang Asli Ethnic Groups in Malaysia. PLoS ONE 7(10): e48165. doi:10.1371/journal.pone.0048165 Editor: Georges Snounou, Universite ´ Pierre et Marie Curie, France Received May 15, 2012; Accepted September 21, 2012; Published October 25, 2012 Copyright: ß 2012 Shahrul Anuar et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by UKMMC Fundamental Research Grant (FF-165-2011), Special Research University Grant (UKM-GUP-2011-316), and Geran Galakan Penyelidik Muda (GGPM-2011-080). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: syahbasree@yahoo.com Introduction Amoebiasis, an infection by the protozoan parasite E. histolytica is globally considered as a leading parasitic cause of human mortality besides malaria and schistosomiasis [1]. It is estimated that E. histolytica may infect half a billion people annually, with 100,000 deaths worldwide [2]. In developed countries, amoebiasis tends to be more common in older individuals and occurs mostly among homosexual men or in institutions [3]. However, in tropical regions, the epidemiology of amoebiasis is completely different and is more common among the general population and particularly among patients attending health care centres with diarrhoea [4]. Clinical features of amoebiasis range from asymptomatic colonization to amoebic dysentery and invasive extraintestinal amoebiasis which is manifested most commonly in the form of liver abscess. Out of 10% of the world’s population infected by E. histolytica, only 1% develops invasive form of the disease [1]. In Malaysia, food and water-borne diseases which are closely associated with environmental and personal hygiene practices are still among the major health problems especially among Orang Asli communities; Malaysian aborigines comprising 0.6% of the total population. The prevalence of E. histolytica/E. dispar/E. moshkovskii in Malaysia ranges from 1% to 61% [5]. To the best of PLOS ONE | www.plosone.org 1 October 2012 | Volume 7 | Issue 10 | e48165