Phylogenetic analysis of simian Plasmodium spp. infecting Anopheles balabacensis Baisas in Sabah, Malaysia.

Tock H Chua,Benny O Manin,Sylvia Daim,Chris Drakeley,Indra Vythilingam,Hans-Peter Fuehrer

doi:10.1371/journal.pntd.0005991

Abstract

BackgroundAnopheles balabacensis of the Leucospyrus group has been confirmed as the primary knowlesi malaria vector in Sabah, Malaysian Borneo for some time now. Presently, knowlesi malaria is the only zoonotic simian malaria in Malaysia with a high prevalence recorded in the states of Sabah and Sarawak.Methodology/Principal findingsAnopheles spp. were sampled using human landing catch (HLC) method at Paradason village in Kudat district of Sabah. The collected Anopheles were identified morphologically and then subjected to total DNA extraction and polymerase chain reaction (PCR) to detect Plasmodium parasites in the mosquitoes. Identification of Plasmodium spp. was confirmed by sequencing the SSU rRNA gene with species specific primers. MEGA4 software was then used to analyse the SSU rRNA sequences and bulid the phylogenetic tree for inferring the relationship between simian malaria parasites in Sabah.PCR results showed that only 1.61% (23/1,425) of the screened An. balabacensis were infected with one or two of the five simian Plasmodium spp. found in Sabah, viz. Plasmodium coatneyi, P. inui, P. fieldi, P. cynomolgi and P. knowlesi. Sequence analysis of SSU rRNA of Plasmodium isolates showed high percentage of identity within the same Plasmodium sp. group. The phylogenetic tree based on the consensus sequences of P. knowlesi showed 99.7%–100.0% nucleotide identity among the isolates from An. balabacensis, human patients and a long-tailed macaque from the same locality.Conclusions/SignificanceThis is the first study showing high molecular identity between the P. knowlesi isolates from An. balabacensis, human patients and a long-tailed macaque in Sabah. The other common simian Plasmodium spp. found in long-tailed macaques and also detected in An. balabacensis were P. coatneyi, P. inui, P. fieldi and P. cynomolgi. The high percentage identity of nucleotide sequences between the P. knowlesi isolates from the long-tailed macaque, An. balabacensis and human patients suggests a close genetic relationship between the parasites from these hosts.

Highlights

Anopheles species of the Leucosphyrus group have been identified as medically important vectors in Southeast Asia region [1,2]
polymerase chain reaction (PCR) results showed that only 1.61% (23/1,425) of the screened An. balabacensis were infected with one or two of the five simian Plasmodium spp. found in Sabah, viz. Plasmodium coatneyi, P. inui, P. fieldi, P. cynomolgi and P. knowlesi
Anopheles balabacensis has been incriminated as the primary vector of zoonotic simian malaria, P. knowlesi in Malaysian Borneo with a high prevalence recorded in the states of Sabah and Sarawak

Summary

Introduction

Anopheles species of the Leucosphyrus group have been identified as medically important vectors in Southeast Asia region [1,2]. Recent studies showed that this species is more active during the early night with a peak biting time between 7 pm to 8 pm [9,13], and prefers to bite outdoors than indoors [13]. Such biting behaviors coupled with an abundant source of simian malaria parasites in the reservoir long-tailed macaques (Macaca fascicularis) contribute to An. balabacensis becoming an effective vector for transmitting P. knowlesi malaria in Sabah.

Methods

Results

Discussion

Conclusion