Factors controlling the long-term temporal and spatial patterns of nitrate-nitrogen export in a dairy farming watershed.

It is difficult to investigate the factors that control the riverine nitrate-nitrogen (NO3--N) export in a watershed which gains or losses groundwater. To control the NO3--N contamination in these watersheds, it is necessary to investigate the factors that are related to the export of NO3--N that is only produced by the watershed itself. This study was conducted in the Shibetsu watershed located in eastern Hokkaido, Japan, which gains external groundwater contribution (EXT) and 34% of the annual NO3--N loading occurs through EXT. The riverine NO3--N exports from 1980 to 2009 were simulated by the SWAT model, and the factors controlling the temporal and spatial patterns of NO3--N exports were investigated without considering the EXT. The results show that hydrological events control NO3--N export at the seasonal scale, while the hydrological and biogeochemical processes are likely to control NO3--N export at the annual scale. There was an integrated effect among the land use, topography, and soil type related to denitrification process, that regulated the spatial patterns of NO3--N export. The spatial distribution of NO3--N export from hydrologic response units (HRUs) identified the agricultural areas with surplus N that are vulnerable to nitrate contamination. A new standard for the N fertilizer application rate including manure application should be given to control riverine NO3--N export. This study demonstrates that applying the SWAT model is an appropriate method to determine the temporal and spatial patterns of NO3--N export from the watershed which includes EXT and to identify the crucial pollution areas within a watershed in which the management practices can be improved to more effectively control NO3--N export to water bodies.

Differences in the Spatial Variability Among CO2, CH 4, and N 2O Gas Fluxes from an Urban Forest Soil in Japan.

The spatial variability of carbon dioxide (CO(2)), methane (CH(4)), and nitrous oxide (N(2)O) fluxes from forest soil with high nitrogen (N) deposition was investigated at a rolling hill region in Japan. Gas fluxes were measured on July 25th and December 5th, 2008 at 100 points within a 100 × 100 m grid. Slope direction and position influenced soil characteristics and site-specific emissions were found. The CO(2) flux showed no topological difference in July, but was significantly lower in December for north-slope with coniferous trees. Spatial dependency of CH(4) fluxes was stronger than that of CO(2) or N(2)O and showed a significantly higher uptake in hill top, and emissions in the valley indicating strong influence of water status. N(2)O fluxes showed no spatial dependency and exhibited high hot spots at different topology in July and December. The high N deposition led to high N(2)O fluxes and emphasized the spatial variability.

Study of the role of anaerobic metabolism in succinate production by Enterobacter aerogenes.

Succinate is a core biochemical building block; optimizing succinate production from biomass by microbial fermentation is a focus of basic and applied biotechnology research. Lowering pH in anaerobic succinate fermentation culture is a cost-effective and environmentally friendly approach to reducing the use of sub-raw materials such as alkali, which are needed for neutralization. To evaluate the potential of bacteria-based succinate fermentation under weak acidic (pH <6.2) and anaerobic conditions, we characterized the anaerobic metabolism of Enterobacter aerogenes AJ110637, which rapidly assimilates glucose at pH 5.0. Based on the profile of anaerobic products, we constructed single-gene knockout mutants to eliminate the main anaerobic metabolic pathways involved in NADH re-oxidation. These single-gene knockout studies showed that the ethanol synthesis pathway serves as the dominant NADH re-oxidation pathway in this organism. To generate a metabolically engineered strain for succinate production, we eliminated ethanol formation and introduced a heterogeneous carboxylation enzyme, yielding E. aerogenes strain ΔadhE/PCK. The strain produced succinate from glucose with a 60.5% yield (grams of succinate produced per gram of glucose consumed) at pH <6.2 and anaerobic conditions. Thus, we showed the potential of bacteria-based succinate fermentation under weak acidic conditions.

Pseudenhygromyxa salsuginis gen. nov., sp. nov., a myxobacterium isolated from an estuarine marsh.

A myxobacterial strain, designated SYR-2(T), was obtained from a mud sample from an estuarine marsh alongside the Yoshino River, Shikoku, Japan. It had rod-shaped vegetative cells and formed bacteriolytic enlarging colonies or so-called 'swarms' in the agar media. Fruiting-body-like globular to polyhedral cell aggregates and myxospore-like spherical to ellipsoidal cells within them were observed. Those features coincided with the general characteristics of myxobacteria. The strain was mesophilic and strictly aerobic. Growth of SYR-2(T) was observed at 18-40 °C (optimum, 30-35 °C), pH 5.5-8.3 (optimum, pH 7.0-7.5) and with 0.0-2.5 % (w/v) NaCl (optimum, 0.2-1.0 %). Both Mg(2+) and Ca(2+) were essential cations for the growth. The predominant fatty acids were iso-C15 : 0 (43.8 %), iso-C17 : 0 (22.4 %) and iso-C16 : 0 (9.6 %). A C20 : 4 fatty acid [arachidonic acid (4.3 %)], iso-C19 : 0 (1.5 %) and anteiso-acids [ai-C15 : 0 (0.5 %), ai-C17 : 0 (0.3 %)] were also detected. The G+C content of the DNA was 69.7 mol%. The strain contained menaquinone-7 (MK-7) as the major respiratory quinone. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain SYR-2(T) belonged to the suborder Nannocystineae, order Myxococcales in the class Deltaproteobacteria, and the strain was most closely related to two type strains of marine myxobacteria, Enhygromyxa salina SHK-1(T) and Plesiocystis pacifica SIR-1(T), with 96.5 % and 96.0 % similarities, respectively. These characteristics determined in this polyphasic study suggested that strain SYR-2(T) represents a novel species in a new genus of myxobacteria. The name Pseudenhygromyxa salsuginis gen. nov., sp. nov. is proposed to accommodate this isolate, and the type strain of Pseudenhygromyxa salsuginis is SYR-2(T) ( = NBRC 104351(T) = DSM 21377(T)).

Assessment of river water quality during snowmelt and base flow periods in two catchment areas with different land use.

River water quality was evaluated with respect to eutrophication and land use during spring snowmelt and summer base flow periods in Abashiri (mixed cropland-livestock farming) and Okoppe (grassland-based dairy cattle farming), eastern Hokkaido, Japan. Water from rivers and tributaries was sampled during snowmelt and summer base flow periods in 2005, and river flow was measured. Total N (TN), NO(3)-N, and Si concentrations were determined using standard methods. Total catchment and upland areas for each sampling site were determined with ArcGIS hydrology modeling software and 1:25,000-scale digital topographic maps. Specific discharge was significantly higher during snowmelt than during base flow. In both areas, TN concentrations increased, whereas Si concentrations decreased, with increased specific discharge, and were significantly higher during snowmelt. The Si:TN mole ratio decreased to below or close to the threshold value for eutrophication (2.7) in one-third of sites during snowmelt. River NO(3)-N concentrations during base flow were significantly and positively correlated with the proportion of upland fields in the catchment in both the Abashiri (r = 0.88, P < 0.001) and Okoppe (r = 0.43, P < 0.01) areas. However, the regression slope, defined as the impact factor (IF) of water quality, was much higher in Abashiri (0.025) than in Okoppe (0.0094). The correlations were also significantly positive during snowmelt in both areas, but IF was four to eight times higher during snowmelt than during base flow. Higher discharge of N from upland fields and grasslands during snowmelt and the resulting eutrophication in estuaries suggest that nutrient discharge during snowmelt should be taken into account when assessing and monitoring the annual loss of nutrients from agricultural fields.