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Viruses belonging to the genus Quaranjavirus in the family Orthomyxoviridae are known as argasid tick-borne viruses. Some viruses in this genus or an unassigned quaranjavirus-like variant can infect humans, although little is known about their pathogenicity. During the surveillance of tick-borne viruses in ixodid ticks in Ehime Prefecture, Japan, novel quaranjavirus-like sequences were detected in 3 pooled samples of Haemaphysalis histricis nymphs. Phylogenetic analysis revealed that the detected viruses formed a cluster with quaranjaviruses and other related viruses. Specifically, the viruses were closely related to Zambezi tick virus 1 and Uumaja virus, which are quaranjavirus-like viruses recently discovered in ixodid ticks in Africa and Europe, respectively. These findings indicate that the viruses detected in this study were probably new members of the Quaranjavirus genus or a related group. The viruses were tentatively named "Ohshima virus" even though only limited sequences of their genomes were available. This is the first report on the detection of a quaranjavirus-like virus in the East Asian region. Further investigations are needed to discern its infectivity and pathogenicity against humans and other animals and to determine the potential risk of an emerging tick-borne viral disease.

Anopheles (Anopheles) lindesayi Giles consists of 5 subspecies. In Japan, only one subspecies, An. l. japonicus Yamada, has been reported. Its geographical populations are morphologically diverse; however, they are regarded as a single subspecies. In this study, we re-evaluated the taxonomic status of An. l. japonicus in Japan, and that of another subspecies, An. l. pleccau, distributed in Taiwan, by comparative morphological and molecular analyses based on the gene sequences of mitochondrial DNA cytochrome c oxidase I (COI) and ribosomal DNA internal transcribed spacer 2 (ITS2). Nucleotide sequence divergence was calculated using the Kimura-two-parameter (K2P) distance model. Phylogenetic trees based on COI and ITS2 sequences showed 3 distinct clades: Eastern Japan, Western Japan, and the Ryukyus. The sequences of the Ryukyu specimens were located within the same clade as that of the sequences of the Taiwanese specimens. Regarding the COI sequences, the 3 geographical groups in Japan were genetically distinct. The following morphological characteristics distinguished the groups: larval seta 1-S, pupal setae 5 through segments IV-VII, and pupal setae 6 on segments IV-VII. Based on these results, it was revealed that An. l. japonicus included 3 genetically and morphologically distinct groups: 2 groups of An. l. japonicus and a group in the Ryukyus, which was a synonym of An. l. pleccau.

Dengue fever, an acute, mosquito-borne, febrile illness caused by Flavivirus spp., is a problem in Africa, South and Southeast Asia, Latin America, and the Caribbean. A dengue outbreak occurred after nearly 70 years of absence or no detection, and then 158 autochthonous cases occurred in Japan from August to October 15, 2014. The most competent mosquito vectors for dengue virus transmission were Aedes aegypti and A. albopictus. Since A. albopictus is widely distributed across Japan and A. aegypti recently invaded Japan by airplane, we examined the susceptibility of these species to infection by dengue virus.

Internal transcribed spacer regions of ribosomal DNA were sequenced, and new species-specific primers were designed to simplify the molecular identification of five morphologically related subgenus Stegomyia mosquito species--Aedes aegypti, Ae. albopictus, Ae. riversi, Ae. flavopictus, and Ae. daitensis--found in the Ryukyu Archipelago, Japan. Each newly designed primer was able to amplify a species-specific fragment with a different size. Conditions for multiplex PCR were optimized to identify all five species in a single PCR. This method is a convenient tool for entomological field surveys, particularly in arbovirus endemic/epidemic areas where some of these species coexist.

A mark-release-recapture study of the dispersal ability of blow flies, Calliphora nigribarbis, was conducted in Ikumo-Naka, Ato Town, Yamaguchi Prefecture, Japan in December 2004. A location where a fatal avian influenza outbreak had occurred 1 year previously was selected for the present study. A total of 3,884 C. nigribarbis were collected, 1,915 of which were marked and released from 4 different collection sites during 2 successive days. The recapture rate of the released C. nigribarbis ranged from 0.014 to 0.029 among the collection sites, and the overall recapture rate was calculated as 0.022. Based on the distance between the released site and the recaptured site, the dispersal rate of C. nigribarbis was estimated as 256 m/h on the 1st day and 179 m/h on the 2nd day of the experiment, and the maximum dispersal rate observed in this study was estimated as 500 m/h. Taking into account the active period of C. nigribarbis on a fine day (7 h/day), the distance traveled by C. nigribarbis within a day was estimated as 1,789 and 1,250 m/day on average for the 1st and 2nd days, respectively, and the maximum distance was 3,500 m/day. These results suggest that C. nigribarbis could play a role in the mechanical dissemination of avian influenza virus and spread of the outbreak in Japan.

The Culex pipiens complex consists of vector mosquitoes that transmit important human pathogens. In this study we established a simplified method to distinguish three members of the Cx. pipiens complex, Cx. p. pallens Coquillet, Cx. p. form molestus Forskal, and Cx. quinquefasciatus Say, collected in Japan. Sequence analysis of the Drosophila Ace-orthologous acetylcholinesterase (Ace) gene (668 to 680 bp) revealed that a single polymorphic region characterizes each species. Based on this region, specific primers that distinguish Cx. p. form molestus (ACEpip2) and Cx. p. pallens (ACEpall2) were newly designed. Polymerase chain reactions were performed with the genomic DNA of Culex mosquitoes as the template, and these primers clearly distinguished two Culex spp. The accuracy of the designed primers was evaluated with 38 colonies of mosquito samples collected from 9 prefectures of Japan. The testing revealed that the distribution of anautogenous Cx. p. pipiens has not been confirmed in Japan. It also revealed that the male of Cx. p. pallens possesses an Ace gene haplotype that is highly similar to the sequence of Cx. quinquefasciatus. This improved method allows the evaluation of vector competence of Cx. p.molestus, which is the suspected vector of West Nile virus.