title: Simian malaria in wild macaques: first report from Hulu Selangor district, Selangor, Malaysia.
abstract: BACKGROUND
  Malaria is a vector-borne parasitic disease which is prevalent in many developing countries. Recently, it has been found that Plasmodium knowlesi, a simian malaria parasite can be life-threatening to humans. Long-tailed macaques, which are widely distributed in Malaysia, are the natural hosts for simian malaria, including P. knowlesi. The aim of the present study was to determine the prevalence of simian malaria parasites in long-tailed macaques in the district of Hulu Selangor, Selangor, Malaysia.
  
  
  METHODS
  A total of 70 blood samples were collected from Macaca fascicularis dwelling in the forest of Hulu Selangor by the Department of Wildlife and National Parks Peninsular Malaysia, Kuala Lumpur, Malaysia. DNA was extracted using PureLink™ Genomic DNA Kits. Conventional and nested PCR were used to detect the genus and species of Plasmodium parasites respectively. In addition, phylogenetic analysis was carried out to confirm the species of Plasmodium parasites.
  
  
  RESULTS
  Thirty-five (50 %) of the 70 samples were positive for Plasmodium using genus-specific primers. These positive samples were then subjected to nested PCR targeting the 18S ribosomal RNA genes to detect all five simian malaria parasites: namely, P. knowlesi, Plasmodium inui, Plasmodium cynomolgi, Plasmodium fieldi, and Plasmodium coatneyi. All five species of simian malaria parasites were detected. Of these, P. inui was the predominant (65.7 %), followed by P. knowlesi (60 %), P. cynomolgi (51.4 %) P. coatneyi (45.7 %) and P. fieldi (2.9 %). A total of nine macaques had mono-infection with P. knowlesi (four), P. cynomolgi (two), P. coatneyi (two) and P. fieldi (one). Eleven of the macaques had dual infections while 12 had triple infections. Three macaques were infected with four species of Plasmodium. Molecular and phylogenetic analysis confirmed the five species of Plasmodium parasites.
  
  
  CONCLUSIONS
  This study has provided evidence to elucidate the presence of transmission of malaria parasites among the local macaques in Hulu Selangor. Since malaria is a zoonosis, it is important to determine the new control strategies for the control of malaria.
authors.affiliation: Department of Medicine, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. rumanahdst@yahoo.com.
authors.forename: Rumana
authors.initials: R
authors.lastname: Akter
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. indra.vythilingam@gmail.com.
authors.forename: Indra
authors.initials: I
authors.lastname: Vythilingam
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. khawloketim@hotmail.com.
authors.forename: Loke Tim
authors.initials: LT
authors.lastname: Khaw
authors.affiliation: Department of Medicine, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. rajesqvist2004@yahoo.com.
authors.forename: Rajes
authors.initials: R
authors.lastname: Qvist
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. limailian@um.edu.my.
authors.forename: Yvonne Ai-Lian
authors.initials: YA
authors.lastname: Lim
authors.affiliation: Department of Wildlife and National Park (PERHILITAN), KM10, Jalan Cheras, 56100, Kuala Lumpur, Malaysia. frankie@wildlife.gov.my.
authors.forename: Frankie Thomas
authors.initials: FT
authors.lastname: Sitam
authors.affiliation: State Vector Borne Disease Control Unit, Selangor State Health Department, Selangor, Malaysia. drbvenu@moh.gov.my.
authors.forename: Balan
authors.initials: B
authors.lastname: Venugopalan
authors.affiliation: Department of Microbiology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. shamalamy@yahoo.com.
authors.forename: Shamala Devi
authors.initials: SD
authors.lastname: Sekaran
mesh_terms: D000818:Animals; D001769:Blood / Q000469:parasitology*; D016000:Cluster Analysis; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D008252:Macaca fascicularis; D008288:Malaria / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*; D008296:Malaysia / Q000453:epidemiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D010961:Plasmodium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D018419:Primate Diseases / Q000175:diagnosis* / Q000453:epidemiology / Q000469:parasitology*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S