著者
Ryoji Suzuki Shouji Houki Kenji Ito Hiroyuki Shibaike
出版者
The Plankton Society of Japan, The Japanese Association of Benthology
雑誌
Plankton and Benthos Research (ISSN:18808247)
巻号頁・発行日
vol.18, no.4, pp.206-213, 2023-11-27 (Released:2023-11-30)
参考文献数
21

An environmental DNA (eDNA) detection technique using TaqMan qPCR and LAMP analysis was developed for the Asian clam Corbicula fluminea, which is causing alarm in Japan and worldwide as an invasive alien species. The qPCR primer and probe designed in this study specifically amplified C. fluminea DNA, but did not amplify C. sandai or C. japonica DNA, both native Japanese species. In contrast, LAMP analysis showed that C. sandai DNA also reacted, but with a delay after the amplification of C. fluminea DNA, and it was possible to specifically detect C. fluminea DNA by setting the LAMP analysis time to 30 min. Field surveys at three study sites with different habitat conditions for C. fluminea showed that eDNA detection using qPCR/LAMP analyses were consistent with C. fluminea distribution. The applicability of a simple filtration and concentration method for eDNA using glass fibers in suspension (SGF method) was verified, and the SGF method had a higher eDNA recovery capacity than the conventional aspiration method using glass fiber filter paper. eDNA extraction using the SGF method and eDNA analysis using qPCR/LAMP may be useful for qualitative and simple habitat estimation of C. fluminea in poor water quality environments.
著者
Kenji Ito Hiroyuki Shibaike
出版者
The Plankton Society of Japan, The Japanese Association of Benthology
雑誌
Plankton and Benthos Research (ISSN:18808247)
巻号頁・発行日
vol.16, no.2, pp.100-108, 2021-05-27 (Released:2021-06-11)
参考文献数
36
被引用文献数
8

The golden mussel Limnoperna fortunei (Dunker, 1857) is an invasive freshwater bivalve species that exerts harmful effects on the environment, as well as man-made structures, such as water-treatment systems. By using conventional sampling methods, it is difficult to detect mussels under low-density conditions; however, environmental DNA (eDNA) analysis may be a rapid and efficient method for monitoring this aquatic organism. In this study, we conducted surveys based on the eDNA analysis of L. fortunei in 15 farm ponds in Japan and compared the results with those of two conventional survey methods, visual census and plankton larval survey, to clarify the effectiveness of eDNA analysis for field surveys of L. fortunei. Primers and a probe specific to L. fortunei were developed, and a method for analysis was established. In the laboratory experiments, the species eDNA was detected in all water tanks containing the mussels, and the concentration of eDNA was high in the experimental tank that had high density of L. fortunei. In the field survey, L. fortunei eDNA was detected in all ponds where the mussels were found by conventional survey, and low concentrations of eDNA were also detected in several ponds where no L. fortunei were found by traditional methods. These results suggest that eDNA analysis has greater sensitivity for the detection of L. fortunei in farm ponds than that of conventional methods. Environmental DNA surveys have little impact on water management and are suitable for surveys at water facilities that have not yet been damaged by the mussels.