Z oonotic malaria caused by Plasmodium knowlesi, commonly found in long-tailed macaques (Macaca fascicularis) and pig-tailed macaques (M. nemestrina), is now a major emerging disease, particularly in Malaysia (1,2). Two other simian malaria parasites, P. cynomolgi (2-4) and P. inui (2), have also been shown to have the potential of zoonotic transmission to humans through the bites of infected mosquitoes under natural and experimental conditions. The risk of acquiring zoonotic malaria is highest for persons living at the forest fringe and working or venturing into the forest because of their proximity with the monkey reservoir hosts and the mosquito vectors (5,6). With the aid of molecular methods, we aimed to investigate whether human infections with simian malaria parasites were present among indigenous communities in Malaysia whose villages are situated in the forest or at the forest fringe. The StudyWe examined 645 archived blood samples that we had collected during 2011-2014 among indigenous populations of various subtribes from 14 villages in 7 states in Malaysia (Appendix Table 1, https://wwwnc. cdc.gov/EID/article/27/8/20-4502-App1.pdf). We fi rst screened the extracted DNA samples at Universiti Malaya (UM) for the presence of Plasmodium with the aid of genus-specifi c primers (rPLU1 and rPLU5; rPLU3 and rPLU4) (Appendix). Of the 645 indigenous community samples, 102 (15.8%) were positive for Plasmodium. Using species-specifi c nested PCR assays (Appendix), we identifi ed these infections as monoinfections with P. knowlesi (n = 40), P. vivax (n = 21), P. cynomolgi (n = 9), P. falciparum (n = 6), P. coatneyi (n = 3), P. inui (n = 3), P. malariae (n = 2), and P. ovale curtisi (n = 1) (Table 1). In 17 samples, the species could not be identifi ed despite repeated attempts. Our species-specifi c primer pairs were designed on the basis of either the asexually (A) or sexually (S) transcribed forms of Plasmodium small subunit (SSU) rRNA genes (7); the genus-specifi c primer pairs anneal to both asexual and sexual forms of the SSU rRNA genes, and therefore the genus-specifi c assay is more sensitive.We further characterized the 55 samples that tested positive for simian malaria parasites by amplifying a longer fragment of the SSU rRNA gene (914 bp-950 bp) for direct sequencing. Phylogenetic analysis using the neighbor-joining method (Figure 1) revealed the presence of P. knowlesi
Background Formerly known as the Malaysian hunter gatherers, the Negrito Orang Asli (OA) were heavily dependent on the forest for sustenance and early studies indicated high prevalence of intestinal parasitism. Initiation of a redevelopment program in the 1970s aimed to demarginalize the OA was expected to reduce soil transmitted helminth (STH) infections. Gradually, the OA were relocated to new resettlement areas at the peripheries. The aim of this study was to compare STH infections between Negritos who are still living in the inland jungle with those living in resettlements. Methodology/Principal findings A total of 416 Negrito participants were grouped into two categories of communities based on location; Inland Jungle Villages (IJV); and Resettlement Plan Scheme (RPS). Iodine wet mount, formalin-ether sedimentation, modified Trichrome and modified Ziehl-Neelsen staining and Kato-Katz methods were performed on stool samples. A questionnaire was used to collect information regarding demographic, socioeconomic, environmental and hygiene behaviors. Prevalence of STH was significantly higher in IJV (91.3%) versus RPS (83.1%) (P = 0.02). However, the percentage of individuals with severe intensity of Trichuris trichiura infections was significantly higher in the RPS (17.2%) compared to IJV (6.5%) (P = 0.01). Severe Ascaris lumbricoides infection was observed at 20.0% amongst RPS Negritos and 15.0% amongst IJV (P = 0.41). Whilst for hookworm infection, both prevalence and individuals with moderate to severe infections were higher in the IJV (26.2%, 41.0%) versus RPS (18.7%, 24.0%) (P values = 0.08, 0.09), accordingly. The prevalence other intestinal parasitic infections (e.g. Entamoeba sp., Blastocystis and flukes) was also higher in IJV versus RPS. Apart from poor hygienic behaviors as significant risk factors in both communities, low socio-economic status was highly associated with STH infections in RPS (P<0.001) but not significantly associated in IJV. Conclusions The findings showed that ex situ development plan by RPS has not profoundly contributed to the STH reduction among the OA. Conversely, burden rate of T . trichiura infections increased due to their extreme poverty and poor hygienic behaviors. Here, we are suggesting biannual mass albendazole intervention (triple dose regimens in RPS, but a single dose in IJV) and community empowerment to both communities. For a long-term and better uptake, these strategies must be done together with the community input and participation, respecting their traditional customs and accompanied by recruitment of more OA people in the health-care taskforce.
Southeast Asia (SEA) is enriched with a complex history of peopling. Malaysia, which is located at the crossroads of SEA, has been recognized as one of the hubs for early human migration. To unravel the genomic complexity of the native inhabitants of Malaysia, we sequenced 12 samples from 3 indigenous populations from Peninsular Malaysia and 4 native populations from North Borneo to a high coverage of 28-37×. We showed that the Negritos from Peninsular Malaysia shared a common ancestor with the East Asians, but exhibited some level of gene flow from South Asia, while the North Borneo populations exhibited closer genetic affinity towards East Asians than the Malays. The analysis of time of divergence suggested that ancestors of Negrito were the earliest settlers in the Malay Peninsula, whom first separated from the Papuans ~ 50-33 thousand years ago (kya), followed by East Asian (~ 40-15 kya), while the divergence time frame between North Borneo and East Asia populations predates the Austronesian expansion period implies a possible pre-Neolithic colonization. Substantial Neanderthal ancestry was confirmed in our genomes, as was observed in other East Asians. However, no significant difference was observed, in terms of the proportion of Denisovan gene flow into these native inhabitants from Malaysia. Judging from the similar amount of introgression in the Southeast Asians and East Asians, our findings suggest that the Denisovan gene flow may have occurred before the divergence of these populations and that the shared similarities are likely an ancestral component.
Despite the tremendous growth of the DNA sequencing data in the last decade, our understanding of the human genome is still in its infancy. To understand the implications of genetic variants in the light of population genetics and molecular evolution, we developed a database, PGG.SNV (https://www.pggsnv.org), which gives much higher weight to previously under-investigated indigenous populations in Asia. PGG.SNV archives 265 million SNVs across 220,147 present-day genomes and 1018 ancient genomes, including 1009 newly sequenced genomes, representing 977 global populations. Moreover, estimation of population genetic diversity and evolutionary parameters is available in PGG.SNV, a unique feature compared with other databases.
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