Bacterial metabolisms influence the behavior of uranium (U) in deep geological repository (DGR) system because bacteria are ubiquitous in the natural environment. Nevertheless, most studies for the U(VI) bioreduction have focused on a few model bacterium, such as Shewanella putrefaciens, Desulfovibrio desulfuricans, and Geobacter sulfurreducens. In this study, the potential of aqueous U(VI) ((U(VI)aq) reduction by indigenous bacteria was examined under anaerobic conditions with addition of 20 mM sodium acetate for 24 weeks. Three different indigenous bacterial communities obtained from granitic groundwater at depths of 44–60 m (S1), 92–116 m (S2), and 234–244 m (S3) were applied for U(VI)aq reduction experiments. The S2 groundwater contained the highest U concentration of 885.4 μg/L among three groundwater samples, where U mainly existed in the form of Ca2UO2(CO3)3(aq). The S2 groundwater amended 20 mM of sodium acetate was used for the U(VI)aq bioreduction experiment. Variations in the U(VI)aq concentration and redox potential were monitored for 24 weeks to compare U(VI)aq removal efficiency in response to indigenous bacteria. The U(VI)aq removal efficiencies varied among three indigenous bacteria: 57.8% (S3), 43.1% (S2), and 37.7% (S1). The presence of the thermodynamically stable uranyl carbonate complex resulted in the incomplete U(VI)aq removal. Significant shifts in indigenous bacterial communities were observed through highthroughput 16S rRNA gene sequencing analysis. Two SRB species, Thermodesulfovibrio yellowstonii and Desulfatirhabdium butyrativorans, were dominant in the S3 sample after the anaerobic reaction, which enhanced the bioreduction of U(VI)aq. The precipitates produced by bacterial activity were determined to be U(IV)-silicate nanoparticles by a transmission electron microscope (TEM)-energy dispersive spectroscope (EDS) analysis. These results demonstrated that considerable U immobilization is possible by stimulating the activity of indigenous bacteria in the DGR environment.

The purpose of this study was to investigate the effects of the concentration of seminal plasma in aerobic and anaerobic conditions on the total motility(TM) and the progressive motility(PM) of spermatozoa in long term preservation of cooled equine semen. We also examine the pregnancy rates after artificial insemination using fresh, cooled or frozen semen, and different durations of cooled-preserved equine semen. In the aerobic state of cooledpreserved semen, As the increase of preserved duration to 24h, 48h, 72h, and 96h, TM tended to decrease in each of different concentrations of formalin-containing experimental group, TM tended to decrease regardless of the concentrations of SP. In different concentrations of SP, TM of without seminal plasma(SP W/O) group tended to be higher than that of SP 20%, SP 33% and SP 50%, especially TM of SP W/O group was significantly higher than other groups at 96 h (p<0.05). PM was higher in the groups of SP W/O and SP 20% than in the groups of SP 33% and SP 50% from 24 h to 72 h in cooled-preservation, especially PM of SP W/O group was significantly higher than other groups at 96 h (p<0.05). In the anaerobic condition of cooled-preserved semen, the results of TM and PM at different concentrations of SP were similar to the results in the aerobic condition although there was a difference in the ratio. The pregnancy rates of fresh-cooled, cooled-preserved and frozen semen were 66.3%, 60.7% and 34.5%, respectively, and the pregnancy rate of frozen semen was the lowest. We also found that it is possible to pregnancy after artificial insemination using 72 h cooled-preserved equine semen. There was similar of the pregnancy rates in the different month from April to August.

Optical microscope, SEM (Scanning Electron Microscopy) and fluorescent microscope were used for qualitative and morphological studies of the attached biomass on PE (polyethylene) substratum under anaerobic condition. It was shown by the observation of optical microscope that the initial attachment of biomass began in crevices of the surface of PE. The shape and structure of the attached biofilm could be observed by SEM photographs, but species of bacteria were and methanogens were not classified. A large number of methanogenic bacteria were identified on the surface of PE substratum by fluorescence under 480nm of radiation. It was estimated that methanogenic bacteria was also related to initial attachment of biomass under anaerobic condition.