ABSTRACT
The Izumi plain in the Kagoshima Prefecture, Japan, is known as an overwintering site for more than 30,000 migratory waterfowl, including endangered crane species. We previously reported that environmental water samples, from artificial wet paddies created as crane roost sites on the Izumi plain, are useful for avian influenza virus (AIV) surveillance. During the 2019/20 winter season, we collected 238 water samples from the crane roost sites and isolated 22 AIVs of six subtypes: one H1N1, one H3N2, seven H3N8, four H4N6, nine H6N6, and one H11N2 subtypes. Genetic analyses revealed that AIVs of the same subtype isolated from the Izumi plain during a single winter season exhibited multiple genetic constellations. Furthermore, phylogenetic analyses suggested that our H3N2 isolate may be a genetic reassortant between close relatives to our H3N8 and H11N2 isolates. Our study highlighted the importance of monitoring AIV circulation to better understand AIV ecology in migratory waterfowl populations.
ABSTRACT
Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease, caused by PRRS virus (PRRSV), that critically affects the swine industry. While the detection of PRRSV genes plays a key role in PRRS control, the PRRSV genome is known to undergo frequent mutation. Nevertheless, primer pairs widely used for the detection of PRRSV genes were designed between 1995 and 2010. The reliability of these primer pairs for the detection of currently circulating PRRSVs is therefore questionable. Here, we investigated the sensitivity of the previously reported primer pairs to detect PRRSV genes that have been recently isolated or detected in Japan. In addition, based on nucleotide sequences from the recent Japanese PRRSVs, we designed four new primer pairs for the detection of PRRSV genes. The sensitivity and specificity of the new primer pairs were evaluated by quantitative reverse transcription PCR using RNA extracted from PRRSV isolates, swine serum, and oral fluid specimens collected from PRRS-affected pigs, and swine sera collected from a PRRSV-free pig farm in Japan. One of novel primer pairs used in our study exhibited greater sensitivity than the previously reported primer pairs, and is thus more reliable for the detection of PRRSV genes.
Subject(s)
Porcine Reproductive and Respiratory Syndrome , Porcine respiratory and reproductive syndrome virus , Animals , Base Sequence , Porcine Reproductive and Respiratory Syndrome/diagnosis , Porcine respiratory and reproductive syndrome virus/genetics , Real-Time Polymerase Chain Reaction , Reproducibility of Results , SwineABSTRACT
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.
