Genetic characterization of bovine respiratory syncytial viruses in Japan between 2017 and 2019.

Bovine respiratory syncytial virus (BRSV) strains that were detected in Kagoshima prefecture and isolated in Hokkaido between 2017 and 2019, together with a BRSV vaccine strain, were subjected to full-genome sequencing. The BRSV strains identified in Japan were found to be genetically close to each other but distant from the vaccine strains. The deduced amino acids at positions 206 and 208 of the glycoprotein (G protein), which form one of the major epitopes of the recent Japanese BRSV strains, were different from those of the vaccine strains. Therefore, the recent Japanese BRSV strains might be antigenically different from the BRSV vaccine strains.

Improved method for avian influenza virus isolation from environmental water samples.

Environmental water-targeted surveillance of migratory aquatic birds at overwintering sites is potentially one of the most effective approaches for understanding the ecology of avian influenza viruses (AIVs). In this study, we improved the method for AIV isolation from environmental water samples by making a minor modification to our previously reported process. We experimentally demonstrated that the AIV recovery efficiency of the modified method was 10-100-fold higher than that of the original method. This improved isolation method allowed us to isolate a considerably larger number of AIV isolates from environmental water samples collected at an overwintering site for tens of thousands of migratory aquatic birds in Japan during the 2018/2019 winter season, compared with those during previous winter seasons. Genetic and phylogenetic analyses revealed that AIVs of the same subtypes with multiple genetic constellations were circulating in a single overwintering site during a single winter season. These findings indicate that our improved isolation method contributes to enhance environmental water-targeted surveillance and to a better understanding of AIV ecology in migratory aquatic bird populations by monitoring ongoing AIV circulation.

Detection and molecular characterization of Babesia sp. in wild boar (Sus scrofa) from western Japan.

Wild animals often act as reservoirs of tick-borne Babesia and Theileria spp., which cause piroplasmosis. Therefore, epidemiological investigations about the distribution of these parasites in wild animals are important for evaluating the transmission risk to humans and livestock. In this study, we surveyed Babesia and Theileria spp. infecting wild boar (Sus scrofa) in Kagoshima and Yamaguchi prefectures and Tsushima island, which are all in western Japan, and performed molecular genetic analyses on the samples. DNA was extracted from either blood or liver samples of wild boar captured in Kagoshima prefecture in 2015, 2016, and 2018 and from blood samples from wild boar captured in Yamaguchi prefecture in 2013-2015 and Tsushima island in 2018. PCR screening for the partial 18S ribosomal RNA gene (18S rRNA) of both Babesia and Theileria spp. in wild boar revealed that 63.9 % (140 of 219 samples) were positive. Sequencing of all positive samples revealed that they were all the same Babesia species. Subsequent phylogenetic analyses showed that the parasite is closely related to Babesia sp. previously detected in the hard tick, Amblyomma testudinarium in Kagoshima, and further analyses suggested that this species is genetically related to Babesia gibsoni. On the other hand, no Theileria were detected in any of the samples. In summary, we observed a high prevalence of B. gibsoni-like Babesia sp. in wild boar in western regions of Japan. The host range, distribution, pathogenicity, and life cycle of this protozoan should be further evaluated.

Nationwide prevalence of Torque teno sus virus 1 and k2a in pig populations in Japan.

Because broad genetic diversity has recently been detected in Torque teno sus viruses (TTSuV1 and TTSuVk2), the viral genome detection method needs to be improved to understand the prevalence of these viruses. Here, we established single PCR-based detection methods for the TTSuV1 and TTSuVk2a genomes with newly designed primer pairs and applied them to investigate the prevalence of TTSuV1 and TTSuVk2a in Japanese pig populations. The results revealed that 98.2% and 81.7% of the pig farms tested positive for the TTSuV1 and TTSuVk2a genomes, respectively, indicating that both TTSuV1 and TTSuVk2a are widespread in Japan.