In October 2011, the National International Health Regulations (IHR) 2005 Focal Point for Malaysia received notification from the United States’ Centers for Disease Control and Prevention (CDC) of a probable Sarcocystis outbreak amongst 23 travellers from six countries who had vacationed on Tioman Island between June and August 2011. The Ministry of Health, Malaysia (MOH) in collaboration with the Department of Veterinary Services, Malaysia (DVS) conducted a cross sectional study in November 2011 to determine the presence of Sarcocystosis among humans, animals and in the environment in Tioman Island. Epidemiological investigations conducted involved a community health survey of 44 residents in Kampung Salang, Tioman and review of outpatient attendance cards for suspected or confirmed cases of Sarcocystosis. Twenty-eight fresh stool samples were collected and sent to the National Public Health Laboratory (NPHL) for detection of Sarcocystis oocysts using fluorescence microscopy. Water samples taken from 27 water sampling points around the island were processed and analysed under the fluorescence microscope using ultraviolet (UV) light at the Institute for Medical Research (IMR) to detect the presence of Sarcocystis sporocyst. DVS collected 84 faecal samples from four types of domesticated animals and then analysed them at the Veterinary Services Centre in Tioman Island for Sarcocystis oocysts and other parasitic ova and cysts using qualitative Floatation Technique. The results showed that Sarcocystis was not present in humans, animals and in the environment in Tioman Island during the study period. Further surveillance among humans, wildlife and the environment is needed to determine Sarcocystis endemicity in Tioman Island.