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Background  Singapore has been certified malaria-free by the World Health Organization since November 1982. However, sporadic autochthonous malaria outbreaks do occur. In one of the most recent outbreaks of vivax malaria, an entomological investigation identified  Anopheles sinensis  as the most probable vector. As metaphase karyotype studies divided  An. sinensis  into two forms, A and B, with different vector competence: the investigation of vector competence of  An. sinensis  found in Singapore was thus pursued using  Plasmodium vivax  field isolates from the Thailand–Myanmar border.    Methods  Adults and larvae  An. sinensis  were collected from Singapore from 14 different locations, using various trapping and collection methods between September 2013 and January 2016. Molecular identification of  An. sinensis  species were conducted by amplifying the  ITS2  and  CO1  region using PCR. Experimental infections of  An. sinensis  using blood from seven patients infected with  P. vivax  from the Thailand–Myanmar border were conducted with  Anopheles cracens  ( An. dirus  B )  as control.    Results  Phylogenetic analysis showed that  An. sinensis  (F 22 , F 2  and collected from outbreak areas) found in Singapore was entirely Form A, and closely related to  An. sinensis  Form A from Thailand. Artificial infection of these Singapore strain  An. sinensis  Form A resulted in the development of oocysts in four experiments, with the number of sporozoites produced by one  An. sinensis  ranging from 4301 to 14,538.    Conclusions  Infection experiments showed that  An. sinensis  Form A from Singapore was susceptible to Thai–Myanmar  P. vivax  strain, suggesting a potential role as a malaria vector in Singapore.

Singapore’s Anopheles sinensis Form A is susceptible to Plasmodium vivax isolates from the western Thailand–Myanmar border