Molecular Identification of Species Belonging to Culex vishnui Subgroup (Diptera: Culicidae), Vectors of Japanese Encephalitis Virus, in Taiwan

Han-Hsuan Chung Center for Diagnostic and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Taiwan;

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Tien-Huang Chen Center for Diagnostic and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Taiwan;

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Pei-Feng Wang Center for Diagnostic and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Taiwan;

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Yoshio Tsuda Department of Medical Entomology, National Institute of Infectious Diseases, Tokyo, Japan

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Hwa-Jen Teng Center for Diagnostic and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Taiwan;

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Shiu-Ling Chen Center for Diagnostic and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Taiwan;

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Classification of mosquitoes with overlapping features remains problematic when using traditional morphological identification alone. In this study, we used molecular methods to elucidate the taxonomic status of Culex tritaeniorhynchus, Culex annulus, and Culex pseudovishnui species as vectors of the Japanese encephalitis virus belonging to the Culex vishnui subgroup and gene flow among them. In this study, 76, 59, and 3 samples of Cx. annulus, Cx. tritaeniorhynchus, and Cx. pseudovishnui, respectively, were collected around Taiwan. Phylogenetic analysis and genetic divergence were based on genomic sequence variations in ribosomal DNA and the internal transcribed spacer (rDNA) and cytochrome c oxidase subunit I (COI). Our results revealed that Cx. annulus and Cx. vishnui are genetically similar and share a gene pool among the species from Taiwan and other Asian countries. However, two hidden taxa of Cx. tritaeniorhynchus, which clustered together according to the rDNA sequences, were discovered based on the COI sequences. In addition, Cx. pseudovishnui has different gene pools from those of the strains from other countries, implying that the population from Taiwan is probably either a unique strain or a sibling species. This study provides molecular information on the taxonomic status of the species in the Cx. vishnui subgroup in Taiwan and gene flow between these species, providing valuable information for vector control operations and the delineation of the evolutionary process.

Author Notes

Financial support: We received financial support from the Taiwan Centers for Disease Control (Grant No. DOH101-DC-2036) and the National Institute of Infectious Diseases of Japan (Grant No. 463-NIID-107). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Current contact information: Han-Hsuan Chung, Tien-Huang Chen, Pei-Feng Wang, Hwa-Jen Teng, and Shiu-Ling Chen, Center for Diagnostic and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Taiwan, E-mails: hhc@cdc.gov.tw, dh1015.chen@gmail.com, freshew@yahoo.com.tw, hjteng101@gmail.com, and sl@cdc.gov.tw. Yoshio Tsuda, Department of Medical Entomology, National Institute of Infectious Diseases, Tokyo, Japan, E-mail: tsuda.yoshio@lecinc.co.jp.

Address correspondence to Shiu-Ling Chen, No.6, Linsen S. Rd., Jhongjheng District, Taipei City 10050, Taiwan. E-mail: sl@cdc.gov.tw
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