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. 2019 Jul 15;12(1):351.
doi: 10.1186/s13071-019-3598-1.

Genetic homogeneity of Anopheles maculatus in Indonesia and origin of a novel species present in Central Java

Triwibowo Ambar Garjito  1   2   3 , Umi Widiastuti  4 , Mujiyono Mujiyono  4 , Mega Tyas Prihatin  4 , Widiarti Widiarti  4 , Riyani Setyaningsih  4 , Siti Alfiah  4 , Barandi Sapta Widartono  5 , Din Syafruddin  6 , Tri Baskoro Tunggul Satoto  7 , Laurent Gavotte  8 , Michael J Bangs  9   10 , Sylvie Manguin  11   12 , Roger Frutos  11   13   14
Affiliations

Genetic homogeneity of Anopheles maculatus in Indonesia and origin of a novel species present in Central Java

Triwibowo Ambar Garjito et al. Parasit Vectors. .

Abstract

Background: Anopheles maculatus (s.s.) is an important vector of malaria in Indonesia. Previously it was considered the only member of the Maculatus Group present in Indonesia. A novel species was recently identified in the Kulon Progo District in Central Java. Until recently, few investigations have been conducted looking at An. maculatus genetic diversity in Indonesia, including allopatric island populations.

Methods: Indonesian An. maculatus (s.l.) samples were collected in several locations in Java, Lesser Sunda Island group, Sumatra and in Kulon Progo (Yogyakarta, central Java) where a novel species has been identified. Samples from a 30-year-old colony of the Kulon Progo population were also included in the analysis. Maximum-likelihood analysis established the phylogenies of the ITS2 (nuclear) and cox1 (mitochondrial) markers. Putative times of separation were based on cox1 genetic distances.

Results: Two species of the Maculatus Group are present in Indonesia. The novel sibling species is more closely related to Anopheles dispar than to An. maculatus (s.s.). Anopheles maculatus (s.s.) samples are homogeneous based on the ITS2 sequences. Indonesian samples and An. dispar belong to the same cox1 maternal lineage and differ from all other known members of the Maculatus Group. Divergence time between the different populations found in Java was estimated using an established cox1 mutation rate.

Conclusions: A novel species within the Maculatus Group, most closely related to An. dispar, is confirmed present in the Kulon Progo area of Central Java. The divergence of this species from An. maculatus (s.s.) is explained by the stable refugia in the Kulon Progo area during the quaternary period of intense volcanic activity throughout most of Java. This novel species awaits detailed morphological description before applying a formal species name. For the interim, it is proposed that the Kulon Progo population be designated An. maculatus var. menoreh to distinguish it from An. maculatus (s.s.).

Keywords: Anopheles maculatus; Indonesia; Maculatus Group; Malaria.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Map of the sampling sites in Indonesia. Each sampling site is indicated as a black spot. The name of sampling sites and samples are indicated. The source of geographical data layers is the Indonesia Geospatial Information Agency who granted the permission and rights to publish this map
Fig. 2
Fig. 2
Phylogenetic analysis of the ITS2 sequences. Kulon Progo samples are identified with blue stars. The phylogenetic tree was constructed using the maximum likelihood (ML) method and the Kimura-2 (K80) evolutionary model in Mega X. To assess the ML tree reliability, bootstraps were tested at 1000 replicates
Fig. 3
Fig. 3
Phylogenetic analysis of the cox1 sequences. a Midpoint rooted tree. b Tree rooted using Anopheles sinensis as outgroup. Kulon Progo samples are identified with stars. Phylogenetic trees were constructed with the maximum likelihood (ML) method and the Kimura-2 (K80) evolutionary model in Mega X. To assess the ML tree reliability, bootstraps were tested with 1000 replicates
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