Abstract
BackgroundThe malaria mosquito Anopheles punctipennis, a widely distributed species in North America, is capable of transmitting human malaria and is actively involved in the transmission of the ungulate malaria parasite Plasmodium odocoilei. However, molecular diagnostic tools based on Internal Transcribed Spacer 2 (ITS2) of ribosomal DNA are lacking for this species. Anopheles punctipennis is a former member of the Anopheles maculipennis complex but its systematic position remains unclear.MethodsIn this study, ITS2 sequences were obtained from 276 An. punctipennis specimens collected in the eastern and midwestern United States and a simple and robust Restriction Fragment Length Polymorphism approach for species identification was developed. The maximum-likelihood phylogenetic tree was constructed based on ITS2 sequences available through this study and from GenBank for 20 species of Anopheles.ResultsThe analysis demonstrated a consistent ITS2 sequence length and showed no indications of intragenomic variation among the samples based on ITS2, suggesting that An. punctipennis represents a single species in the studied geographic locations. In this study, An. punctipennis was found in urban, rural, and forest settings, suggesting its potential broad role in pathogen transmission. Phylogeny based on ITS2 sequence comparison demonstrated the close relationship of this species with other members of the Maculipennis group.ConclusionsThis study developed molecular tools based on ITS2 sequences for the malaria vector An. punctipennis and clarified the phylogenetic position of the species within the Maculipennis group.
Highlights
The malaria mosquito Anopheles punctipennis, a widely distributed species in North America, is capable of transmitting human malaria and is actively involved in the transmission of the ungulate malaria parasite Plasmodium odocoilei
Internal Transcribed Spacer 2 sequencing The primary goal of this study was to develop a molecular tool based on ITS2 for the identification of An. punctipennis
A molecular diagnostic tool based on ITS2 sequences, the most reliable species identification tool for malaria mosquitoes [15], had never been developed for An. punctipennis prior to current investigation
Summary
The malaria mosquito Anopheles punctipennis, a widely distributed species in North America, is capable of transmitting human malaria and is actively involved in the transmission of the ungulate malaria parasite Plasmodium odocoilei. In the USA, numerous outbreaks of malaria occurred from as far north as Massachusetts to subtropical southern states in the eighteenth and nineteenth centuries. Each year, the Center for Disease Control and Prevention (CDC) reports between 1,000 and 1,500 cases of malaria in the USA [4], with 63 outbreaks of locally transmitted malaria occurring since 1957 [5]. Human migration, political instability, and the presence of competent malaria vectors increase the risk of malaria and other tropical diseases being imported and transmitted in regions where they were eradicated [5]
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