title: Molecular evidence of Sarcocystis nesbitti in water samples of Tioman Island, Malaysia.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
  
  
  
  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.
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia.
authors.forename: Shahhaziq
authors.initials: S
authors.lastname: Shahari
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia.
authors.forename: Tengku Idzzan Nadzirah
authors.initials: TI
authors.lastname: Tengku-Idris
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia.
authors.forename: Mun Yik
authors.initials: MY
authors.lastname: Fong
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. lauyeeling@um.edu.my.
authors.forename: Yee Ling
authors.initials: YL
authors.lastname: Lau
mesh_terms: D016000:Cluster Analysis; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D062312:Islands; D008296:Malaysia; D008853:Microscopy; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012522:Sarcocystis / Q000145:classification / Q000166:cytology / Q000235:genetics / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA; D014867:Water / Q000469:parasitology*
chemical_list: D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S; D014867:Water
keywords: Sarcocystis; Sarcocystis nesbitti; Sporocysts; Surveillance study; Tioman Island; Water sample