Kazuyo Nagaosa

Unique distribution of deep groundwater bacteria constrained by geological setting

We collected groundwater samples at depths of up to 482 m from three boreholes drilled into sedimentary rock within two formations in Hokkaido, Japan. The prokaryotic community in each subsurface groundwater sample was analysed by microscopic counts and cloning-sequencing the 16S rRNA genes. On total direct counts, there were between 4.61 × 10(4) and 5.06 × 10(6) prokaryote cells ml(-1) in the samples, which is similar to the numbers observed at the marine subsurface. However, the vertical distribution of the prokaryotes did not show a simple decrease in abundance with increasing depth. A high abundance of cells with significant amounts of RNA was identified in the domain Bacteria using fluorescence in situ hybridization, with a high frequency of dividing cells at the transition zone between the two sedimentary rock formations. Cloning-sequencing analysis showed the predominance of γ-Proteobacteria at this transition zone at 281-312 m. The horizontal heterogeneity of the microbial distribution in the subsurface environment was also demonstrated by a relatively high density of members of the domain Archaea in borehole HDB-4, drilled only 1.5 km northeast of HDB-6 and in the same formation.

Subsurface bacterial growth and grazing impact revealed by an in situ experiment in a shallow aquifer

To elucidate bacterial population dynamics in an aquifer, we attempted to reveal the impact of protozoan grazing on bacterial productivity and community structure by an in situ incubation experiment using a diffusion chamber. The abundance and vertical distribution of bacteria and protozoa in the aquifer were revealed using wells that were drilled in a sedimentary rock system in Itako, Ibaraki, Japan. The water column in the wells possessed aerobic and anaerobic layers. Active bacterial populations under the grazing pressure of protozoa were revealed through in situ incubation with grazer eliminating experiment by the filtration. On August 19, 2003, the total number of bacteria (TDC) decreased from 1.5 × 106 cells ml− 1 at 2.2 m depth to 3.0 × 105 cells ml− 1 at 10 m depth. The relative contribution of the domain Bacteria to TDC ranged between 63% and 84%. Protozoa existed at a density of 4.2 × 104 to 1.9 × 105 cells ml− 1 in both aerobic and microaerobic conditions. A grazing elimination experiment in situ for 6 days brought about clearly different bacterial community profiles between the 2.2 m and 10 m samples. The bacterial composition of the initial community was predominantly β- and γ -proteobacteria at 2.2 m, while at 10 m β-, α - and γ -proteobacteria represented 56%, 26% and 13% of the community, respectively. The distribution of bacterial abundance, community composition and growth rates in the subsurface were influenced by grazing as well as by geochemical factors (dissolved oxygen and concentrations of organic carbon, methane and sulfate). Results of the in situ incubation experiment suggested that protozoan grazing contributes significantly to bacterial population dynamics.

Microbes in Groundwater of a Volcanic Mountain, Mt. Fuji; 16S rDNA Phylogenetic Analysis as a Possible Indicator for the Transport Routes of Groundwater

Huge groundwater reserves are under the foot of Mt. Fuji, the tallest volcanic mountain in Japan, and the residence time of the groundwater as estimated from the 36Cl/Cl isotope ratio is in the range of 20–35 years. All springwater samples contain nearly saturated concentrations of oxygen. The groundwater field was separated into four areas based on regional slopes of the groundwater table and estimated groundwater flow directions. Springwater samples showed that physicochemical characteristics of the groundwater in the region are relatively constant; Ion concentrations were characterized as the Ca-HCO3 type, irrespective of location at the foot of the mountain. The density of prokaryotes in the springwater, which was in the range of 102–104 cells mL−1, was low compared with any other groundwater so far reported. Although the density of prokaryotes was low, retrieved clones show that the prokaryotes belonged to eight bacterial and two archaeal phyla, and included both aerobic and obligate anaerobic prokaryotes. Some retrieved clones were related to thermophilic prokaryotes, which are optimally adapted to temperatures greater than 40°C. This finding suggests that at least some of the source of the groundwater was located at a depth of 600 m or greater, based on a temperature gradient of 4°C/100 m. This depth is far below the lava layer which was taken to be a substantial pool of groundwater. Microbiological information may provide insight into the transport route of groundwater until it emerges.