title: Non-invasive surveillance for Plasmodium in reservoir macaque species.
abstract: BACKGROUND
  Primates are important reservoirs for human diseases, but their infection status and disease dynamics are difficult to track in the wild. Within the last decade, a macaque malaria, Plasmodium knowlesi, has caused disease in hundreds of humans in Southeast Asia. In order to track cases and understand zoonotic risk, it is imperative to be able to quantify infection status in reservoir macaque species. In this study, protocols for the collection of non-invasive samples and isolation of malaria parasites from naturally infected macaques are optimized.
  
  
  METHODS
  Paired faecal and blood samples from 60 Macaca fascicularis and four Macaca nemestrina were collected. All animals came from Sumatra or Java and were housed in semi-captive breeding colonies around West Java. DNA was extracted from samples using a modified protocol. Nested polymerase chain reactions (PCR) were run to detect Plasmodium using primers targeting mitochondrial DNA. Sensitivity of screening faecal samples for Plasmodium was compared to other studies using Kruskal Wallis tests and logistic regression models.
  
  
  RESULTS
  The best primer set was 96.7 % (95 % confidence intervals (CI): 83.3-99.4 %) sensitive for detecting Plasmodium in faecal samples of naturally infected macaques (n = 30). This is the first study to produce definitive estimates of Plasmodium sensitivity and specificity in faecal samples from naturally infected hosts. The sensitivity was significantly higher than some other studies involving wild primates.
  
  
  CONCLUSIONS
  Faecal samples can be used for detection of malaria infection in field surveys of macaques, even when there are no parasites visible in thin blood smears. Repeating samples from individuals will improve inferences of the epidemiology of malaria in wild primates.
authors.affiliation: Eijkman Institute for Molecular Biology, Jakarta, Indonesia. pipin@eijkman.go.id.
authors.forename: Josephine E
authors.initials: JE
authors.lastname: Siregar
authors.affiliation: Department of Ecology and Evolutionary Biology, Princeton University, Princeton, NJ, 08544, USA. cfaust@princeton.edu.
authors.forename: Christina L
authors.initials: CL
authors.lastname: Faust
authors.affiliation: Eijkman Institute for Molecular Biology, Jakarta, Indonesia. lydia@eijkman.go.id.
authors.forename: Lydia S
authors.initials: LS
authors.lastname: Murdiyarso
authors.affiliation: Pusat Studi Satwa Primata, Institut Pertanian Bogor, Bogor, Indonesia. lisrosmanah@gmail.com.
authors.forename: Lis
authors.initials: L
authors.lastname: Rosmanah
authors.affiliation: Pusat Studi Satwa Primata, Institut Pertanian Bogor, Bogor, Indonesia. uussaepuloh@yahoo.com.
authors.forename: Uus
authors.initials: U
authors.lastname: Saepuloh
authors.affiliation: Department of Ecology and Evolutionary Biology, Princeton University, Princeton, NJ, 08544, USA. dobber@princeton.edu.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Dobson
authors.affiliation: Pusat Studi Satwa Primata, Institut Pertanian Bogor, Bogor, Indonesia. atie@indo.net.id.
authors.forename: Diah
authors.initials: D
authors.lastname: Iskandriati
mesh_terms: D000818:Animals; D001769:Blood / Q000469:parasitology*; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D004197:Disease Reservoirs; D062665:Epidemiological Monitoring; D005243:Feces / Q000469:parasitology*; D007214:Indonesia / Q000453:epidemiology; D008252:Macaca fascicularis / Q000469:parasitology; D008254:Macaca nemestrina / Q000469:parasitology; D008288:Malaria / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*; D008992:Monkey Diseases / Q000453:epidemiology / Q000469:parasitology*; D016790:Plasmodium knowlesi / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D016054:DNA, Protozoan