Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

In recent year, marine safety has been one of top concerns in Korea. In this paper, general statistics of ship in 159 waterways in Korea from 2007 to 2017 were considered. Main objective of this research is to investigate the relationship between the number of marine accidents and traffic conditions in narrow waterways by multiple regression analysis method. The result shows that the number of vessels, the width, the length and the depth of narrow waterway have an influence on the number of maritime accidents in corresponding area. Additionally, the number of vessels sailing has the most significant impact on the number of accidents in narrow waterway area.

Lead leached out from the metal lead-sinker attached to fishing nets or contained in ropes are likely to cause the environmental contamination, consequently, resulting in ecotoxicity. However, it is very difficult to find out the data on the disposal reality of fishing nets, lead-containing form of/in fishing net and its content. In previous studies, we presented the containing form of lead and the lead concentration leached out by the containing forms. As a result, the leaching concentration of lead was the highest under the supposition condition of lead-sinker deposited on sandy tidal flat. In this study, as a subsequent study, we investigated long-term leaching characteristics from lead-sinker deposited on sandy tidal flat. We used a serial batch leaching experiment to investigate the long-term leaching characteristics, and the experimental conditions were as follows; solid to liquid ratio 1:20, shaking time 24 hours (repetition of 7 times), room temperature, and shaking rate 170 rpm. As a result, lead leaching concentration after shaking for 24 hours was 0.215 mg/L, showing the highest leaching concentration, and then the leaching concentrations between 2-7 times were greatly decreased. Analysis result by Visual Minteq 3.1 said most of the leached lead component could be extracted as crystal such as Anglesite (PbSO4), Cerrusite (PbCO3), Cotunnite (PbCl2), and Pb(OH)2 by sulfate and carbonate in seawater.