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Background    Sarcocystis  are intracellular protozoan parasites that are characterised by their ability to invade muscle tissue and form intramuscular sarcocysts. A muscular sarcocystosis outbreak was reported by travellers returning from Tioman Island in 2011 and 2012 where  Sarcocystis nesbitti  was identified as the main cause. The source of the  S. nesbitti  that was involved has remained elusive, although water is hypothesised to be the main cause of transmission. A surveillance study was therefore undertaken in the northern regions of Tioman Island to identify the source of  S. nesbitti  by screening rivers, water tanks, wells and seawater.    Methods  Water samples were collected from rivers, water tanks, wells and seawater on Tioman Island over the course of April to October 2015. Water samples were indirectly screened for  Sarcocystis  species by obtaining sediment from respective water sources. PCR amplification of the 18S rRNA gene region was conducted to identify positive samples. Microscopy was used in an attempt to reappraise PCR results, but no sporocysts were detected in any of the samples.    Results  A total of 157 water samples were obtained and 19 were positive for various  Sarcocystis  species. Through BLASTn and phylogenetic analysis, these species were found to be  S. singaporensis ,  S. nesbitti ,  Sarcocystis  sp. YLL-2013 and one unidentified  Sarcocystis  species.    Conclusions  This is the first positive finding of  S. nesbitti  in water samples on Tioman Island, which was found in a water tank and in river water samples. This finding supports the hypothesis that water was a potential medium for the transmission of  S. nesbitti  during the outbreak. This will potentially identify areas in which preventive measures can be taken to prevent future outbreaks.

Molecular evidence of Sarcocystis nesbitti in water samples of Tioman Island, Malaysia