RT Journal Article SR Electronic T1 Identification of fish spawning based on size-selected sampling and diurnal concentration changes of environmental DNA resulting from sperm-release JF bioRxiv FD Cold Spring Harbor Laboratory SP 2023.06.19.545639 DO 10.1101/2023.06.19.545639 A1 Tsuji, Satsuki A1 Shibata, Naoki YR 2023 UL http://biorxiv.org/content/early/2023/06/22/2023.06.19.545639.abstract AB Environmental DNA (eDNA) analysis is a useful method for detecting spawning behaviour by observing elevated eDNA concentrations and/or nuclear/mitochondrial DNA ratios. However, little is known about the characteristics and dynamics of sperm-derived eDNA, which is key to detection. This study focused on changes in eDNA particle size distribution (PSD) and concentrations pre-, post-, and during spawning of Plecoglossus altivelis, to investigate the potential for semi-selective recovery of sperm-derived eDNA and the importance of sampling time for improving detection sensitivity. Firstly, PSD changes between pre- and post-spawning were investigated by comparing concentrations and proportions of eDNA obtained from filters with different pore sizes. Secondly, the diurnal changes in eDNA concentration were monitored at the peak of the spawning season by collecting river water every 1-3 hours. Results showed that eDNA related to sperm-head size increased at post-spawning, and eDNA concentrations had significant diurnal changes with a peak during the spawning time window. These findings suggest that semi-selective recovery of sperm-derived eDNA based on particle size and/or sampling during the spawning time window with increased concentrations can improve the detection sensitivity of eDNA-based spawning surveys. This study provides essential fundamental information for advancing eDNA-based spawning surveys and contributes to their further development.Competing Interest StatementThe authors have declared no competing interest.