Cyanobacterial resting cells, such as akinetes, are important seed cells for cyanobacteria’s early development and bloom. Due to their importance, various methods have been attempted to isolate resting cells present in the sediment. Ludox is a solution mainly used for cell separation in marine sediments, but finding an accurate method for use in freshwater is difficult. This study compared the two most commonly used Ludox methods (direct sediment treatment and sediment distilled water suspension treatment). Furthermore, we proposed a highly efficient method for isolating cyanobacterial resting cells and eDNA amplification from freshwater sediments. Most of the resting cells found in the sediment were akinete to the Nostocale and were similar to those of Dolichospermum, Cylindrospermum, and Aphanizomenon. Twenty times more akinetes were found in the conical tube column using the sediment that had no treatment than in the sample treated by suspending the sediment in distilled water. Akinete separated through Ludox were mainly spread over the upper and lower layers in the column rather than concentrated at a specific depth in the column layer. The mibC, Geo, and 16S rDNA genes were successfully amplified using the sediment directly in the sample. However, the amplification products of all genes were not found in the sample in which the sediment was suspended in distilled water. Therefore, 5 g to 10 g of sediment is used without pretreatment when isolating cyanobacterial resting cells from freshwater sediment. Cell isolation and gene amplification efficiency are high when four times the volume of Ludox is added. The Ludox treatment method presented in this study isolates cyanobacterial resting cells in freshwater sediment, and the same efficiency may not appear in other biotas. Therefore, to apply Ludox to the separation of other biotas, it is necessary to conduct a pre-experiment to determine the sediment pretreatment method and the water layer where the target organism exists.

Environmental DNA (eDNA) can exist in both intracellular and extracellular forms in natural ecosystems. When targeting harmful cyanobacteria, extracellular eDNA indicates the presence of traces of cyanobacteria, while intracellular eDNA indicates the potential for cyanobacteria to occur. However, identifying the “actual” potential for harmful cyanobacteria to occur is difficult using the existing sediment eDNA analysis method, which uses silica beads and cannot distinguish between these two forms of eDNA. This study analyzes the applicability of a density gradient centrifugation method (Ludox method) that can selectively analyze intracellular eDNA in sediment to overcome the limitations of conventional sediment eDNA analysis. PCR was used to amplify the extracted eDNA based on the two different methods, and the relative amount of gene amplification was compared using electrophoresis and Image J application. While the conventional bead beating method uses sediment as it is to extract eDNA, it is unknown whether the mic gene amplified from eDNA exists in the cyanobacterial cell or only outside of the cell. However, since the Ludox method concentrates the intracellular eDNA of the sediment through filtration and density gradient, only the mic gene present in the cyanobacteria cells could be amplified. Furthermore, the bead beating method can analyze up to 1 g of sediment at a time, whereas the Ludox method can analyze 5 g to 30 g at a time. This gram of sediments makes it possible to search for even a small amount of mic gene that cannot be searched by conventional bead beating method. In this study, the Ludox method secured sufficient intracellular gene concentration and clearly distinguished intracellular and extracellular eDNA, enabling more accurate and detailed potential analysis. By using the Ludox method for environmental RNA expression and next-generation sequencing (NGS) of harmful cyanobacteria in the sediment, it will be possible to analyze the potential more realistically.

This experiment was conducted to confirm the possibility of preparing Sorghum×sudangrass hybrid artificial hay using far-infrared rays in Korea. The machine used in this experiment is a drying device based on far-infrared rays, and is designed to control temperature, air flow rate, far-infrared radiation amount, and air flow speed. The Sorghum×sudangrass hybrids harvested in late September were wilted in the field for one day, and a drying test was performed on them. Conditions for drying were performed by selecting a total of 7 conditions, and each condition induced a change in radiation amount in a single condition (42%) and two steps (4 treatments) and three steps (2 treatments). The speed of the air flow in the device was fixed at 60 m/s, and the run time was changed to 30, 60, and 90 minutes. The average dry matter (DM) content was 82.84%. The DM content was 59.94 and 76.91%, respectively, in drying conditions 1 and 3, which were not suitable for hay. In terms of drying rate, it was significantly higher than 80% in the 5, 6 and 7 treatment, and power consumption was slightly high with an average of 5.7 kw/h. As for the feed value according to each drying condition, the crude protein (CP) content increased as the drying time increased, and there was no significant difference between treatments in ADF, NDF, IVDMD and TDN content. In terms of RFV, treatment 1, which is a single condition, was significantly lower than the complex condition. Through the above results, it was determined that the drying conditions 4 and 5 were the most advantageous when considering the drying speed, power consumption, and quality.

This study analyzes the importance and implementation of countermeasures to reduce safety accidents that may occur during fishing in coastal gillnet fishing. Safety accidents that may occur in coastal gillnet fishing were classified into 56 measurement items in six areas: slips and trips, contact with machinery, falls, struck by object, musculoskeletal symptoms, and electrical shock. 35 fishermen belonging to coastal gillnet associations in Tongyeong and Namhae were selected as samples, and the importance and implementation of the proposed measurement items were investigated in the field. In addition, the results were evaluated visually by IPA analysis. As a result of the analysis, the importance of struck by object was the highest, and the implementation of slips and trips was the highest. On the other hand, prevention of musculoskeletal damage was found to be the lowest in both importance and implementation. The area with the most significant difference in importance and implementation by the classified area for measurement items was struck by object, and the area with the minor difference was slips and trips. Among the measurement items, the importance is highly recognized, but there are items with low implementation. To improve their level of implementation, countermeasures such as continuous education, economic support, and structural change of fishing boats should be prepared; moreover, policy support activities for implementation should be carried out.