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The Okinawa Trough (OT) is a back-arc basin with a wide distribution of active cold seep systems. However, our understanding of the metabolic function of microbial communities in the cold seep sediments of the OT remains limited. In this study, we investigated the vertical profiles of functional genes involved in methane, nitrogen, and sulphur cycling in the cold seep sediments of the OT. Furthermore, we explored the possible coupling mechanisms between these biogeochemical cycles. The study revealed that the majority of genes associated with the nitrogen and sulphur cycles were most abundant in the surface sediment layers. However, only the key genes responsible for sulphur disproportionation ( sor ), nitrogen fixation ( nifDKH ), and methane metabolism ( mcrABG ) were more prevalent within sulfate-methane transition zone (SMTZ). Significant positive correlations ( P < 0.05) were observed between functional genes involved in sulphur oxidation, thiosulphate disproportionation with denitrification, and dissimilatory nitrate reduction to ammonium (DNRA), as well as between AOM/methanogenesis and nitrogen fixation, and between sulphur disproportionation and AOM. A genome of Filomicrobium (class Alphaproteobacteria) has demonstrated potential in chemoautotrophic activities, particularly in coupling DNRA and denitrification with sulphur oxidation. Additionally, the characterized sulfate reducers such as Syntrophobacterales have been found to be capable of utilizing nitrate as an electron acceptor. The predominant methanogenic/methanotrophic groups in the OT sediments were identified as H 2 -dependent methylotrophic methanogens (Methanomassiliicoccales and Methanofastidiosales) and ANME-1a. This study offered a thorough understanding of microbial ecosystems in the OT cold seep sediments, emphasizing their contribution to nutrient cycling.IMPORTANCEThe Okinawa Trough (OT) is a back-arc basin formed by extension within the continental lithosphere behind the Ryukyu Trench arc system. Cold seeps are widespread in the OT. While some studies have explored microbial communities in OT cold seep sediments, their metabolic potential remains largely unknown. In this study, we used metagenomic analysis to enhance comprehension of the microbial community's role in nutrient cycling and proposed hypotheses on the coupling process and mechanisms involved in biogeochemical cycles. It was revealed that multiple metabolic pathways can be performed by a single organism or microbes that interact with each other to carry out various biogeochemical cycling. This data set provided a genomic road map on microbial nutrient cycling in OT sediment microbial communities., Competing Interests: The authors declare no conflict of interest.

Nitrogen (N) resorption from senescing leaves enables plants to reuse N, thereby making them less dependent on current N uptake from the environment. Therefore, N resorption is important for survival and fitness, particularly for plants growing under low N supply. We studied N resorption from senescing leaves of 25 legumes and 25 nonlegumes in a temperate region of Japan to test the hypothesis that high N resorption has not evolved in legumes that fix atmospheric N 2 . The extent of N resorption was quantified by N resorption proficiency that is measured as the level to which leaf N concentration was reduced during senescence, i.e., the lower the senesced leaf N concentration, the lower the N loss through leaf fall and higher the N resorption proficiency. In support of the hypothesis, senesced leaf N concentration was higher in legumes than in nonlegumes, but there was considerable overlap between the groups. The higher senesced leaf N concentration of legumes was associated with a lower proportion of leaf N resorbed during senescence, particularly in species with higher leaf N concentrations. According to a hierarchical partitioning analysis, there was a large contribution of species to the total variance in the senesced leaf N concentration as opposed to a minor contribution of functional group (legume/nonlegume). This study reveals that legumes are not proficient at resorbing N from senescing leaves but that N 2 -fixation might not be the single most important determinant of N resorption.

The increasing eutrophication of freshwater and brackish habitats globally has led to a corresponding increase in the occurrence of harmful cyanobacterial blooms. Cyanobacteria can produce highly toxic substances such as microcystins (MCs) that affect the health of livestock, wildlife, and humans. The present study broaden the understanding of cyanobacteria ecology and MC dynamics in the field, focusing on the estimation of the production and sedimentation rates of MCs in a natural habitat. The nutrient concentrations of the reservoir water and sediment pore water were monitored at 3-h intervals for 24 h during the summer cyanobacterial bloom. The DIN uptake rate of Microcystis in the Isahaya reservoir was estimated and the large-scale blooms in the reservoir were largely controlled by the interactions between rainfall and nutrient levels in the warm season. By using calculations based on the nitrogen budgets and tracking changes of the MC concentrations in the water column, the total MC production and sedimentation rates were estimated to be 52.2 kg MCs d -1 and 21.5 kg MCs d -1 , respectively. Although MCs could be degraded in the environment, the MC sedimentation still comprised 41% of the in-water production., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Recent advances in molecular and genetic studies about flowering time control have been increasingly available to elucidate the physiological mechanism underlying masting, the intermittent and synchronized production of a large amount of flowers and seeds in plant populations. To identify unexplored developmental and physiological processes associated with masting, genome-wide transcriptome analysis is a promising tool, but such analyses have yet to be performed. We established a field transcriptome using a typical masting species, Japanese beech (Fagus crenata Blume), over two years, and analyzed the data using a nonlinear time-series analysis called convergent cross mapping. Our field transcriptome was found to undergo numerous changes depending on the status of floral induction and season. An integrated approach of high-throughput transcriptomics and causal inference was successful at detecting novel causal regulatory relationships between nitrate transport and florigen synthesis/transport in a forest tree species. The synergistic activation of nitrate transport and floral transition could be adaptive to simultaneously satisfy floral transition at the appropriate timing and the nitrogen demand needed for flower formation.

In mountainous areas, plant distribution is constrained by various environmental stresses. Plasticity and constancy in plant functional traits may relate to optimal strategies at respective habitats and to ecotypic differentiation along elevation. Although plant biomass allocation has been extensively studied in relation to adaptation to soil nutrient availability along elevation, its optimality is still poorly understood. We examined soil nutrient availability in the field and conducted growth analysis for two elevational ecotypes of Arabidopsis halleri grown under different nutrient availabilities. We determined plasticity in morphological and physiological traits and evaluated optimal biomass allocation using an optimality model. Our field investigation indicated that soil nitrogen (N) availability increased rather than decreased with increasing elevation. Our growth analysis revealed that lowland ecotype was more plastic in morphological variables and N concentrations, whereas the highland ecotype was more plastic in other physiological variables such as the net assimilation rate (NAR). The leaf mass ratio (LMR) in the lowland ecotype was moderately plastic at the whole range of N availabilities, whereas LMR in the highland ecotype was very plastic at higher N availabilities only. The optimality model indicated that the LMR of the lowland ecotype was nearly optimal throughout the range of studied N availabilities, whereas that of the highland ecotype was suboptimal at low N availability. These results suggest that highland ecotype is adapted only to high N availability, whereas the lowland ecotype is adapted to a relatively wide range of N availabilities as a result of natural selection in their respective habitats. We conclude that an adaptive differentiation has occurred between the two ecotypes and plasticity in the biomass allocation is directly related to its optimization in changing environments.

Tricholoma matsutake is an ectomycorrhizal (ECM) fungus capable of in vitro saprotrophic growth, but the sources of C and N used to generate sporocarps in vivo are not well understood. We examined natural abundance isotope data to investigate this phenomenon. For this purpose, C, N and their stable isotopes ( 13 C, 15 N) content of fungal sporocarps and their potential nutrient sources (i.e., foliage, litter, fine roots, wood, and soil) were investigated from two well-studied sites in Finland and Japan. Our results show that δ 13 C values of T. matsutake and other fungal groups are consistent with those of most studies, but a very high δ 15 N value (16.8‰ ± 2.3) is observed in T. matsutake. Such isotopic pattern of fungal δ 15 N suggests that matsutake has a greater proteolytic potential to digest chemically complex 15 N-enriched organic matter and hydrophobic hyphae. This assumption is further supported by a significant and positive correlation between δ 13 C cap-stipe and δ 15 N cap-stipe exclusively in T. matsutake, which suggests common C and N sources (protein) possible for isotopically enriched cap. The 13 C increase of caps relative to stipe presumably reflects greater contents of 13 C-enriched protein than 13 C-depleted chitin. We conclude that T. matsutake is a typical ECM fungus which obtains for its sporocarp development for both C and N from a common protein source (vs. photosynthetic carbon) present in soil organic matter.

We estimated the nitrogen (N), phosphorus (P), potassium (K) and magnesium (Mg) excretion factors of broiler and layer chickens in Japan, using two approaches and the latest data available. In the top-down approach, we determined the nutrient amounts in the feeds and those in the products (i.e. the liveweight gain, eggs), and the national nutrient excretions were determined as the difference between these amounts. We then calculated the nutrient excretion factors by dividing the national excretions by the number of animals. In the bottom-up approach, we calculated the amounts of nutrients in the feed and product per head using productivity parameters (feed conversion ratio, etc.). The differences between these amounts were considered the nutrient excretion factors. The average nutrient excretion factors of broilers (g/day/head) estimated using the top-down and bottom-up approaches were: N, 1.40 and 1.87; P, 0.36, 0.50; K, 0.54, 0.77; Mg, 0.13, 0.18, respectively. The excretion factors obtained by the top-down approach can be used to calculate the national/regional excretions. The two approaches resulted in almost the same excretion factors for layers, and the average nutrient excretion factors of layers (g/day/head) estimated were: N, 2.20; P, 0.55; K, 0.68; Mg, 0.23. The estimated excretion factors for N (only) are smaller than the reported factors. [ABSTRACT FROM AUTHOR]

The juice from sweet sorghum cultivar SIL-05 (harvested at physiological maturity) was extracted, and the component sucrose and reducing sugars (such as glucose and fructose) were subjected to a membrane separation process to purify the sucrose for subsequent sugar refining and to obtain a feedstock for repeated bioethanol production. Nanofiltration (NF) of an ultrafiltration (UF) permeate using an NTR-7450 membrane (Nitto Denko Corporation, Osaka, Japan) concentrated the juice and produced a sucrose-rich fraction (143.2 g L -1 sucrose, 8.5 g L -1 glucose, and 4.5 g L -1 fructose). In addition, the above NF permeate was concentrated using an ESNA3 NF membrane to provide concentrated permeated sugars (227.9 g L -1 ) and capture various amino acids in the juice, enabling subsequent ethanol fermentation without the addition of an exogenous nitrogen source. Sequential batch fermentation using the ESNA3 membrane concentrate provided an ethanol titer and theoretical ethanol yield of 102.5-109.5 g L -1 and 84.4-89.6%, respectively, throughout the five-cycle batch fermentation by Saccharomyces cerevisiae BY4741. Our results demonstrate that a membrane process using UF and two types of NF membranes has the potential to allow sucrose purification and repeated bioethanol production.

Long-term monitoring of ecosystem succession provides baseline data for conservation and management, as well as for understanding the dynamics of underlying biogeochemical processes. We examined the effects of deforestation and subsequent afforestation of a riparian forest of Japanese cedar (Cryptomeria japonica) on stable isotope ratios of carbon (δ¹³C) and nitrogen (δ¹⁵N) and natural abundances of radiocarbon (Δ¹⁴C) in stream biota in the Mt. Gomadan Experimental Forest and the Wakayama Forest Research Station, Kyoto University, central Japan. Macroinvertebrates, periphytic algae attached to rock surfaces (periphyton), and leaf litter of terrestrial plants were collected from six headwater streams with similar climate, topography, and bedrock geology, except for the stand ages of riparian forests (from 3 to 49 yr old in five stands and > 90 yr old in one reference stand). Light intensity and δ¹³C values of both periphyton and macroinvertebrates decreased synchronously with forest age in winter. A Bayesian mixing model indicates that periphyton contributions to the stream food webs are maximized in 23-yr-old forests. Except for grazers, most macroinvertebrates showed Δ¹⁴C values similar to those of terrestrial leaf litter, reflecting the influence of modern atmospheric CO₂ Δ¹⁴C values. On the other hand, the Δ¹⁴C values of both periphyton and grazers (i.e., aquatic primary consumers) were significantly lower than that of modern atmospheric CO₂, and were lowest in 23-yr-old forest stands. Previous studies show that root biomass of C. japonica peaks at 15-30 yr after planting. These evidences suggest that soil CO₂ released by root respiration and dispersed by groundwater weathers carbonate substrata, and that dissolved inorganic carbon (DIC) with low Δ¹⁴C is incorporated into stream periphyton and some macroinvertebrates. The ecological response in the studied streams to clear-cutting and replanting of Japanese cedar is much slower (~20 yr) than the chemical response (< 5 yr). More than 50 yr is required for the food web structure to completely recover from clear-cutting. The ecological delay is attributed to several biogeochemical factors, the understanding of which is critical to integrated management of forest-stream continuum and the prediction of ecosystem resilience in response to environmental change.

Premise of the Study: Differences in leaf size are expected to be coordinated with various shoot traits and branching intensity because these relationships will influence light capture efficiency, water use, and biomechanics. Previous studies have mainly focused on interspecific patterns of these trait relationships, but not on intraspecific patterns at the geographic scale. We investigated intraspecific variation in shoot traits and branching intensity of Fagus crenata in Japan., Methods: Allometric relationships between the traits of current-year shoots and branching intensity per branch unit of 1-m length on the main axis (BI) and its coordination with latitude were investigated using trees from 10 provenances in a common garden., Key Results: Individual trees originating from lower latitudes have smaller leaves with greater leaf mass per area and nitrogen content per area, greater Huber value (stem cross-sectional area per total leaf area [ATL]) of current-year shoots, and greater BI. Notably, the slope of the log-log relationship between BI and ATL was close to -1.0 across the trees from different source sites, implying that branching in this species occurs to control leaf area., Conclusions: Shoot traits and branching intensity were apparently coordinated with leaf size to control leaf area deployment in this species. Such patterns probably reflect differences in competition for hydraulic conductance among nearby shoots within crowns, as a consequence of different meteorological conditions across the source sites., (© 2015 Botanical Society of America, Inc.)

Nitrogen (N) digestion and urea-N metabolism in Hokkaido native horses fed roughage-based diets containing different types and levels of protein sources were studied. Horses (173 ± 4.8 kg) fitted with an ileum cannula were fed four diets consisting of 100% timothy hay (TH), 88% TH and 12% soybean meal (SBM), 79% TH and 21% SBM, and 51% TH and 49% alfalfa hay at 2.2% of body weight. Dietary protein content varied from 5% to 15% of dry matter. Apparent N digestibilities in the pre-cecum and total tract for the TH diet were lower than those for other diets. However, the proportion of post-ileum N digestion to N intake was not affected by the diets. Urea-N production was linearly related to N intake, but gut urea-N entry was not affected by the diets. The proportion of gut urea-N entry to urea-N production tended to be higher for the TH diet (57%) than the two SBM diets (39%). Anabolic use of urea-N entering the gut was not affected by the diets (20-36% of gut urea-N entry). These results indicate that urea-N recycling provides additional N sources for microbial fermentation in the hindgut of Hokkaido native horses fed low-quality roughages., (© 2014 Japanese Society of Animal Science.)

Microalgal storage lipids are considered to be a promising source for next-generation biofuel feedstock. However, microalgal biodiesel is not yet economically feasible due to the high cost of production. One of the reasons for this is that the use of a low-cost open pond system is currently limited because of the unavoidable contamination with undesirable organisms. Extremophiles have an advantage in culturing in an open pond system because they grow in extreme environments toxic to other organisms. In this study, we isolated the acidophilic green alga Pseudochlorella sp. YKT1 from sulfuric acid mine drainage in Nagano Prefecture, Japan. The vegetative cells of YKT1 display the morphological characteristics of Trebouxiophyceae and molecular phylogenetic analyses indicated it to be most closely related to Pseudochlorella pringsheimii. The optimal pH and temperature for the growth of YKT1 are pH 3.0-5.0 and a temperature 20-25°C, respectively. Further, YKT1 is able to grow at pH 2.0 and at 32°C, which corresponds to the usual water temperature in the outdoors in summer in many countries. YKT1 accumulates a large amount of storage lipids (∼30% of dry weigh) under a nitrogen-depleted condition at low-pH (pH 3.0). These results show that acidophilic green algae will be useful for industrial applications by acidic open culture systems.

The development of crops which are well suited to growth under future environmental conditions such as higher atmospheric CO2 concentrations ([CO2]) is essential to meeting the challenge of ensuring food security in the face of the growing human population and changing climate. A high-yielding indica rice variety (Oryza sativa L. cv. Takanari) has been recently identified as a potential candidate for such breeding, due to its high productivity in present [CO2]. To test if it could further increase its productivity under elevated [CO2] (eCO2), Takanari was grown in the paddy field under season-long free-air CO2 enrichment (FACE, approximately 200 µmol mol(-1) above ambient [CO2]) and its leaf physiology was compared with the representative japonica variety 'Koshihikari'. Takanari showed consistently higher midday photosynthesis and stomatal conductance than Koshihikari under both ambient and FACE growth conditions over 2 years. Maximum ribulose-1,5-bisphosphate carboxylation and electron transport rates were higher for Takanari at the mid-grain filling stage in both years. Mesophyll conductance was higher in Takanari than in Koshihikari at the late grain-filling stage. In contrast to Koshihikari, Takanari grown under FACE conditions showed no decrease in total leaf nitrogen on an area basis relative to ambient-grown plants. Chl content was higher in Takanari than in Koshihikari at the same leaf nitrogen level. These results indicate that Takanari maintains its superiority over Koshihikari in regards to its leaf-level productivity when grown in elevated [CO2] and it may be a valuable resource for rice breeding programs which seek to increase crop productivity under current and future [CO2].

Based on quantification and qualification of bacterial 16S rDNA, we verified the bacterial ecological characteristics of surface sediments of Lakes Shinji and Nakaumi, which are representative of coastal lagoons in Japan. Quantification and qualification of the 16S rDNA sequences was carried out using real time polymerase chain reaction and polymerase chain reaction denaturing gradient gel electrophoresis and non-metric multidimensional scaling, respectively. The results revealed that the copy number per gram of sediment ranged from 8.33 × 10(8) (Lake Nakaumi) to 1.69 × 10(11) (Honjo area), suggesting that bacterial carbon contributed only 0.05-9.64 % of the total carbon content in the samples. Compared with other aquatic environments, these results indicate that sedimentary bacteria are not likely to be important transporters of nutrients to higher trophic levels, or to act as carbon sinks in the lagoons. The bacterial compositions of Lake Shinji and Lake Nakaumi and the Honjo area were primarily influenced by sediment grain sizes and salinity, respectively. Statistical comparisons of the environmental properties suggested that the areas that were oxygen-abundant (Lake Shinji) and at a higher temperature (Honjo area) presented efficient organic matter degradation. The 16S rDNA copy number per gram of carbon and nitrogen showed the same tendency. Consequently, the primary roles of bacteria were degradation and preservation of organic materials, and this was affected by oxygen and temperature. These roles were supported by the bacterial diversity rather than the differences in the community compositions of the sedimentary bacteria in these coastal lagoons.

We set up a free-air ozone (O(3)) exposure system for determining the photosynthetic responses of Siebold's beech (Fagus crenata) and oak (Quercus mongolica var. crispula) to O(3) under field conditions. Ten-year-old saplings of beech and oak were exposed to an elevated O(3) concentration (60 nmol mol(-1)) during daytime from 6 August to 11 November 2011. Ozone significantly reduced the net photosynthetic rate in leaves of both species in October, by 46% for beech and 15% for oak. In beech there were significant decreases in maximum rate of carboxylation, maximum rate of electron transport in photosynthesis, nitrogen content and photosynthetic nitrogen use efficiency, but not in oak. Stomatal limitation of photosynthesis was unaffected by O(3). We therefore concluded photosynthesis in beech is more sensitive to O(3) than that in oak, and the O(3)-induced reduction of photosynthetic activity in beech was due not to stomatal closure, but to biochemical limitation., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Objectives: To estimate protein requirements in older hospitalized adults with pressure ulcers (PrU) according to systemic conditions and wound severity., Design: Secondary nitrogen balance study over 3 days., Setting: Long-term care facility., Participants: Twenty-eight older adults with PrU using a urinary catheter., Measurements: Nitrogen balance over 3 days was evaluated from habitual nitrogen intake measured using a food weighing record and nitrogen excretion from urine, feces and wound exudate. Nitrogen intake required to maintain nitrogen equilibrium was estimated as an average protein requirement using a linear mixed model., Results: Nitrogen intake at nitrogen equilibrium was 0.151 gN/kg per day (95% confidence interval = 0.127-0.175 gN/kg per day) for all participants. The amount of protein loss from wound exudate contributed little to total nitrogen excretion. A Charlson comorbidity index of 4 or greater (the median value) was related to lower nitrogen intake at nitrogen equilibrium (P = .005). Severe PrU with heavy exudate amounts and measured wound areas of 7.9 cm(2) or greater (the median value) were related to higher nitrogen intake at nitrogen equilibrium in individuals with a Charlson comorbidity index of 3 or less (both P = .04). Larger wound area (correlation coefficient (r) = 0.55, P = .003) and heavier exudate volume (r = 0.53, P = .004) were associated with muscle protein hypercatabolism measured according to 3-methylhistidine/creatinine ratio., Conclusion: The average protein requirement is 0.95 g/kg per day for older hospitalized Japanese adults with PrU, but protein requirements depend on an individual's condition and wound severity and range from 0.75 to 1.30 g/kg per day. Severe PrU can require higher protein intakes because of muscle protein hypercatabolism rather than direct loss of protein from wound exudate., (© 2012, Copyright the Authors Journal compilation © 2012, The American Geriatrics Society.)

This study investigated the fleshy fruit characteristics of 28 woody species in a Japanese temperate forest where large sedentary seed-dispersing mammals are present. We tested whether the findings in previous studies in temperate forests of Europe and North America are universal or not. Results have suggested that fruits of all species were eaten both by birds and mammals except for four species with larger fruits, which were eaten only by mammals. A gradient was found from a syndrome characterized by small, oily, and large-seeded fruits to a syndrome characterized by large, succulent, non-oily, and small-seeded fruits. The sizes and colors of the fruits were not conspicuously different from previous findings in Europe and North America. On the other hand, nitrogen and lipids in the fleshy part did not show seasonally increasing trends, or even seasonally decreasing trends in terms of dry weight. This result, suggesting the absence of community-level adaptation of fruit traits to migratory bird dispersers, contrasted with findings in Europe and North America. Large sedentary arboreal or tree-climbing mammals may have a greater effect on the evolution of fruit-disperser relations than opportunistic migratory birds.

Japanese paddy rice systems commonly adopt the rotation of vegetables, wheat and soybeans with paddy rice. Crop rotation may, however, increase the nutrient load in effluent discharged from the district because more fertilizer is applied to the rotation crops than is applied to paddy crops. We investigated a paddy-field district subject to collective crop rotation and quantified the annual nutrient load of effluent from the district in three consecutive years. The total annual exports of nitrogen and phosphorus over the investigation period ranged from 30.3 to 40.6 kg N ha(-1) and 2.62 to 3.13 kg P ha(-1). The results suggest that rotation cropping increases the effluent nutrient load because applied fertilizer is converted to nitrate, and surface runoff is increased due to the absence of shuttering boards at the field outlets.

The hybrid larch F(1) (Larix gmelinii var. japonica × Larix kaempferi) is considered one of the most important tree species not only for timber production but also as an afforestation material for severe conditions such as infertile soil. To predict the ability of hybrid larch F(1) as an afforestation material under potential climates in the future, it is important to understand the response of hybrid larch F(1) to elevated CO(2) concentration ([CO(2)]) under low nutrient availability. Three-year-old seedlings of hybrid larch F(1) were grown under two different levels of [CO(2)], 360 (ambient) and 720 µmol mol(-1) (elevated), in combination with two different levels of nitrogen (N) supply (0 and 30 kg ha(-1)) for one growing season. Elevated [CO(2)] reduced the maximum rates of carboxylation and electron transport in the needles. Net photosynthetic rates at growth [CO(2)] (i.e., 360 and 720 µmol mol(-1) for ambient and elevated treatment, respectively) did not differ between the two CO(2) treatments. Reductions in N content and N use efficiency to perform photosynthetic functions owing to the deficiency of nutrients other than N, such as P and K, and/or increase in cell wall mass were considered factors of photosynthetic down-regulation under elevated [CO(2)], whereas stomatal closure little affected the photosynthetic down-regulation. Although we observed strong down-regulation of photosynthesis, the dry matter increase of hybrid larch F(1) seedlings was enhanced under elevated [CO(2)]. This is mainly attributable to the increase in the amount of needles on increasing the number of sylleptic branches. These results suggest that elevated CO(2) may increase the growth of hybrid larch F(1) even under low nutrient availability, and that this increase may be regulated by changes in both crown architecture and needle photosynthesis, which is mainly affected not by stomatal limitation but by biochemical limitation.

A coccoid- and amorphous-shaped, non-gliding, reddish bacterium, designated SG-29(T), was isolated from seawater in the western North Pacific Ocean near Japan. The strain was Gram-stain-negative, obligately aerobic, heterotrophic and catalase-positive. Nitrate was reduced to nitrogen and acid was produced from aesculin, turanose, 2-keto-gluconate and arabinose. Growth occurred with 1-5 % NaCl (optimum 2 % NaCl) and at 5-37 °C (optimum 20-30 °C). The G+C content of genomic DNA was 68.9 mol% (HPLC). The predominant isoprenoid quinone was MK-7 and the major cellular fatty acids (>10 %) were iso-C₁₇:₁ω9c, C₁₇:₁ω8c and iso-C₁₇:₀. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SG-29(T) belonged to the phylum Bacteroidetes. The most closely related strain was Rhodothermus obamensis JCM 9785(T), with which the isolate exhibited 86.1 % 16S rRNA gene sequence similarity. On the basis of phenotypic and phylogenetic features, strain SG-29(T) is classified in a novel genus and species within the family 'Rhodothermaceae', for which the name Rubricoccus marinus gen. nov., sp. nov. ( = NBRC 107124(T)  = KCTC 23197(T)) is proposed.

Hydraulic limitations associated with increasing tree height result in reduced foliar stomatal conductance (g(s)) and light-saturated photosynthesis (A(max)). However, it is unclear whether the decline in A(max) is attributable to height-related modifications in foliar nitrogen concentration (N), to mesophyll conductance (g(m)) or to biochemical capacity for photosynthesis (maximum rate of carboxylation, V(cmax)). Simultaneous measurements of gas exchange and chlorophyll fluorescence were made to determine g(m) and V(cmax) in four height classes of Pinus densiflora Sieb. & Zucc. trees. As the average height of growing trees increased from 3.1 to 13.7 m, g(m) decreased from 0.250 to 0.107 mol m(-2) s(-1), and the CO(2) concentration from the intercellular space (C(i)) to the site of carboxylation (C(c)) decreased by an average of 74 µmol mol(-1). Furthermore, V(cmax) estimated from C(c) increased from 68.4 to 112.0 µmol m(-2) s(-1) with the increase in height, but did not change when it was calculated based on C(i). In contrast, A(max) decreased from 14.17 to 10.73 µmol m(-2) s(-1). Leaf dry mass per unit area (LMA) increased significantly with tree height as well as N on both a dry mass and an area basis. All of these parameters were significantly correlated with tree height. In addition, g(m) was closely correlated with LMA and g(s), indicating that increased diffusive resistance for CO(2) may be the inevitable consequence of morphological adaptation. Foliar N per unit area was positively correlated with V(cmax) based on C(c) but negatively with A(max), suggesting that enhancement of photosynthetic capacity is achieved by allocating more N to foliage in order to minimize the declines in A(max). Increases in the N cost associated with carbon gain because of the limited water available to taller trees lead to a trade-off between water use efficiency and photosynthetic nitrogen use efficiency. In conclusion, the height-related decrease in photosynthetic performance appears to result mainly from diffusive resistances rather than biochemical limitations.

An indica variety Takanari is known as one of the most productive rice varieties in Japan and consistently produces 20-30% heavier dry matter during ripening than Japanese commercial varieties in the field. The higher rate of photosynthesis of individual leaves during ripening has been recognized in Takanari. By using pot-grown plants under conditions of minimal mutual shading, it was confirmed that the higher rate of leaf photosynthesis is responsible for the higher dry matter production after heading in Takanari as compared with a japonica variety, Koshihikari. The rate of leaf photosynthesis and shoot dry weight became larger in Takanari after the panicle formation and heading stages, respectively, than in Koshihikari. Roots grew rapidly in the panicle formation stage until heading in Takanari compared with Koshihikari. The higher rate of leaf photosynthesis in Takanari resulted not only from the higher content of leaf nitrogen, which was caused by its elevated capacity for nitrogen accumulation, but also from higher stomatal conductance. When measured under light-saturated conditions, stomatal conductance was already decreased due to the reduction in leaf water potential in Koshihikari even under conditions of a relatively small difference in leaf-air vapour pressure difference. In contrast, the higher stomatal conductance was supported by the maintenance of higher leaf water potential through the higher hydraulic conductance in Takanari with the larger area of root surface. However, no increase in root hydraulic conductivity was expected in Takanari. The larger root surface area of Takanari might be a target trait in future rice breeding for increasing dry matter production.

To develop cleaved amplified polymorphic sequence (CAPS) markers for cultivar identification of the tea leaf, 5 primer pairs designed on the basis of genes that encode proteins related to nitrogen assimilation and 26 primer pairs based on expressed sequence tag (EST) sequences of the root of tea plant were screened. From combinations of primer pair and restriction enzyme that showed polymorphism among tea plants, 16 markers were selected and applied to DNA fingerprinting of Japanese tea cultivars. Sixty-three cultivars, except for a bud sport (Kiraka) and its original cultivar (Yabukita) and a pair that was the progeny of the same crossing parent (Harumoegi and Sakimidori), were distinguished from one another. By combining the 16 markers with previously developed CAPS markers and observing the physical appearance, 67 cultivars were distinguishable. The cultivars involve approximately 95% of total tea cultivating area in Japan; therefore, about 95% of tea leaves produced in Japan can be authenticated by labeling their cultivars.

Nitrogen is generally the most limiting nutrient for rice production. In rice paddy soils, various biochemical processes can occur regarding N cycling, including nitrification, denitrification, and nitrogen fixation. Since its discovery in the 1930s, the nitrification-denitrification process has been extensively studied in Japan. It may cause N loss from rice paddy soils, while it can also reduce environmental pollutions such as nitrate leaching and emission of nitrous oxide (N(2)O). In this review article, we first summarize the early and important findings regarding nitrification-denitrification in rice paddy soils, and then update recent findings regarding key players in denitrification and N(2)O reduction. In addition, we also discuss the potential occurrence of other newly found reactions in the N cycle, such as archaeal ammonia oxidization, fungal denitrification, anaerobic methane oxidation coupled with denitrification, and anaerobic ammonium oxidation.

Photosynthetic capacity is known to vary considerably among species. Its physiological cause and ecological significance have been one of the most fundamental questions in plant ecophysiology. We studied the contents of Rubisco (a key enzyme of photosynthesis) and cell walls in leaves of 26 species with a large variation in photosynthetic rates. We focused on photosynthetic nitrogen-use efficiency (PNUE, photosynthetic rate per nitrogen), which can be expressed as the product of Rubisco-use efficiency (RBUE, photosynthetic rate per Rubisco) and Rubisco nitrogen fraction (RNF, Rubisco nitrogen per total leaf nitrogen). RBUE accounted for 70% of the interspecific variation in PNUE. The variation in RBUE was ascribed partly to stomatal conductance, and other factors such as mesophyll conductance and Rubisco kinetics might also be involved. RNF was also significantly related to PNUE but the correlation was relatively weak. Cell wall nitrogen fraction (WNF, cell wall nitrogen per total leaf nitrogen) increased with increasing leaf mass per area, but there was no correlation between RNF and WNF. These results suggest that nitrogen allocation to cell walls does not explain the variation in PNUE. The difference in PNUE was not caused by a sole factor that was markedly different among species but by several factors each of which was slightly disadvantageous in low PNUE species.

A flocculent yeast, Hansenula anomala J224 PAWA, bred in this study, accumulated twice as much phosphorus as the wild type. Over a 30-d period, PAWA removed 70-80% of dissolved total phosphorus from sweet-potato and barley shochu wastewaters (alcoholic distillery wastewaters) while the wild type removed only 30%. Waste sludge was easily separated from effluent wastewater because PAWA cells made large flocks that rapidly settled. Component analysis suggested that PAWA sludge could be used as a protein source for feedstuff and as a phosphorus source for fertilizer. Under anaerobic conditions, denitrification was rapid, resulting in the removal of large amounts of nitrogen from barley shochu wastewater. These results suggest that small shochu manufacturers could benefit from using PAWA to remove phosphorus and organic compounds and then by using a combination of the upflow anaerobic sludge blanket and the downflow hanging sponge method (UASB-DHS method) for nitrification/denitrification.

During the summers (July and August) of 2002-2005, we measured interannual variation in maximum carboxylation rate (V(cmax)) within a Fagus crenata Blume crown in relation to climate variables such as air temperature, daytime vapor pressure deficit (VPD) and daily photosynthetic photon flux, leaf nitrogen per unit area (N(a)) and leaf mass per unit area (LMA). Climatic conditions in the summers of 2002-2004 differed markedly, with warm and dry atmospheric conditions in 2002, cool, humid and cloudy conditions in 2003, and warm clear conditions in 2004. Conditions in summer 2005 were intermediate between those of summers 2002 and 2003, and similar to recent (8-year) means. In July, marked interannual variation in V(cmax) was mainly observed in leaves in the high-light environment (relative photon flux > 50%) within the crown. At the crown top, V(cmax) was about twofold higher in 2002 than in 2003, and V(cmax) values in 2004 and 2005 were intermediate between those in 2002 and 2003. In August, although interannual variation in V(cmax) among the years 2003, 2004 and 2005 was less, marked variation between 2002 and the other study years was evident. Multiple regression analysis of V(cmax) against the climate variables revealed that VPD of the previous 10-30 days had a significant influence on variability in V(cmax). Neither N(a), LMA nor leaf CO(2) conductance from the stomata to the carboxylation site explained the variability in V(cmax). Our results indicate that the long-term climatic response of V(cmax) should be considered when estimating forest carbon gain across the year.

The expression of Japanese chestnut (Castanea crenata) agglutinin (CCA) and its mRNA was investigated in nitrogen-fertilized young potted plants and in floral organs of adult trees. Two levels of N were used: 10 and 20mM NH(4)NO(3). Both levels increased protein content in all vegetative organs, though the magnitude of the increase differed. The highest increase was observed in stems. High levels were retained in 20mM N-fertilized plants, whereas the protein content decreased at 10mM N fertilization. Expression of CCA and its mRNA was observed in young leaves and stems, and their quantities depended on the amount of N fertilizer supplied. In mature leaves, CCA was detected in the first 4 weeks, but its mRNA was undetectable throughout the experimental period. Neither CCA nor its mRNA was detected in roots. In floral organs, CCA and its mRNA were expressed throughout the flower but their quantities differed. These results suggest that CCA acts as a vegetative storage protein, which functions in temporary nitrogen reserve. The results also suggest that expression of CCA is regulated at both transcriptional and translational levels.

Denitrification (as N(2) flux) and organic matter mineralization (as O(2) uptake) were simultaneously measured in the same set of core sediments from a natural sandy and a constructed muddy tidal flat of Ago Bay, Japan. Denitrification rates at both tidal flats fluctuated between ca. 2-20 micromol N(2)m(-2)h(-1) without showing a clear seasonal pattern, and appeared to be substrate limited as NO(3)(-) enrichment (final concentration ca. 225 microM) caused prompt and similar enhancements of ca. 10-folds. Organic matter mineralization rates were markedly higher at constructed muddy flat compared to those of natural sandy flat, especially in summer, and exhibited pronounced temperature dependent (p<0.01) seasonality for both tidal flats. O(2) uptake rates were generally ca. 2-3 order greater than respective denitrification rates indicating dominance of mineralization processes over N(2) losses.

Nitrogen budgets and predominant benthic organisms were examined at a matured artificial tidal flat of Osaka Nanko bird sanctuary and an artificial young tidal flat of Hannan Second District, Osaka Bay, Japan. At the Osaka Nanko bird sanctuary, indexes of the quality of the bottom sediment increased over the time course and no abiotic area was observed in the macrobenthic fauna. Dissolved inorganic nitrogen imported into the tidal flat changed its form and was exported in the form of dissolved organic nitrogen. As a result, total nitrogen was trapped at the tidal flat at an average rate of 138 mgN m(-2)day(-1). At the young tidal flat of Hannan Second District net exchange of nitrogen varied markedly among 2000, 2001, 2002 and 2003. In September 2000 when only 5 months had passed after construction, the tidal flat functioned as a site of source for nitrogen. However, it changed as a sink for nitrogen in 2001-2002 in relation to the growth of seaweeds (Ulva spp. and Gracilaria vermiculophylla) and clams (Tapes philippinarum). In 2003 nitrogen was again released from the artificial tidal flat due to the decrease of biomass of the dominant organisms. Comparing the nitrogen budget between the two sites on a per unit time and area basis, net exchanges of nitrogen by tidal exchange were apparently low and relatively constant at the tidal flat of Osaka Nanko bird sanctuary. This suggests that the aged artificial tidal flat of Osaka Nanko bird sanctuary has a stable ecosystem compared to the young artificial tidal flat of Hannan Second District.

Growth characteristics of Picea glehnii Masters, P. jezoensis (Sieb. et Zucc) Carr., P. jezoensis var. hondoensis (Mayr) Rehder and P. shirasawae Hayashi from Japan, P. abies (L.) Karst. from Europe and P. glauca Voss, P. mariana Britt., Sterns and Pogg. and P. rubens Sarg. from North America were compared. The trees were grown in similar conditions at the Tomakomai Experimental Forest of Hokkaido University in northern Japan. Tree growth, needle biomass, longevity, photosynthetic rate, nitrogen concentration and specific leaf area (SLA) were measured, and photosynthetic nitrogen-use efficiency was calculated. Picea jezoensis, P. jezoensis var. hondoensis, P. abies and P. glauca had high growth rates, high photosynthetic rates in young needles, high needle nitrogen concentrations and short needle life spans. In contrast, P. glehnii, P. shirasawae, P. mariana and P. rubens had low growth and photosynthetic rates, low needle nitrogen concentrations, long needle life spans and maintained a high photosynthetic nitrogen-use efficiency in older needles. Examination of relationships between several growth parameters of the eight taxa revealed positive correlations between SLA and mass-based photosynthetic rate and between SLA and mass-based nitrogen concentration, whereas mass-based photosynthetic rate and mass-based nitrogen concentration were negatively correlated with needle longevity. The species differed greatly in growth characteristics despite being grown in similar conditions.

A sulfate-reducing enrichment culture originating from coastal marine sediment of the eutrophic Tokyo Bay, Japan, was successfully established with Casamino acids as a substrate. A thiosulfate reducer, strain C/G2(T), was isolated from the enrichment culture after further enrichment with glutamate. Cells of strain C/G2(T) were non-motile rods (0.6-0.8 microm x 2.2-4.8 microm) and were found singly or in pairs and sometimes in short chains. Spores were not formed. Cells of strain C/G2(T) stained Gram-negatively, despite possessing Gram-positive cell walls. The optimum temperature for growth was 28-30 degrees C, the optimum pH was around 7.8 and the optimum salt concentration was 20-30 g l(-1). Lactate, pyruvate, serine, cysteine, threonine, glutamate, histidine, lysine, arginine, Casamino acids, peptone and yeast extract were fermented as single substrates and no sugar was used as a fermentative substrate. A Stickland reaction was observed with some pairs of amino acids. Fumarate, alanine, proline, phenylalanine, tryptophan, glutamine and aspartate were utilized only in the presence of thiosulfate. Strain C/G2(T) fermented glutamate to H2, CO2, acetate and propionate. Thiosulfate and elemental sulfur were reduced to sulfide. Sulfate, sulfite and nitrate were not utilized as electron acceptors. The growth of strain C/G2(T) on Casamino acids or glutamate was enhanced by co-culturing with Desulfovibrio sp. isolated from the original mixed culture enriched with Casamino acids. The DNA G+C content of strain C/G2(T) was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C/G2(T) formed a distinct cluster with species of the genus Sedimentibacter. The closest relative was Sedimentibacter hydroxybenzoicus (with a gene sequence similarity of 91 %). On the basis of its phylogenetic and phenotypic properties, strain C/G2(T) (=JCM 13356(T)=NBRC 101112(T)=DSM 17477(T)) is proposed as representing a new genus and novel species, Dethiosulfatibacter aminovorans gen. nov., sp. nov.

Recycling of kitchen garbage is an urgent task for reducing public spending and environmental burdens by incineration and/or landfill. There is an interesting regional effort in Ogawa, Saitama prefecture, Japan, in which source-separated kitchen garbage is anaerobically fermented with a biogas plant and the resultant effluent is used as a quick-release organic fertilizer by surrounding farmers. However, scientific assessments of fertilizer values and risks in the use of the effluent were lacking. Thus, a field experiment was conducted from 2003 to 2004 in Tohoku National Agricultural Research Center to grow spinach (Spinacia oleracea L.) and komatsuna (Brassica rapa var. perviridis L. H. Bailey) for evaluating the fertilizer value of the kitchen garbage effluent (KGE), nitrate, coliform group (CG), Escherichia coli, fecal streptococci (FS), and Vibrio parahaemolyticus concentrations of KGE and in the soil and the plant leaves. A cattle manure effluent (CME) and chemical fertilizers (NPK) were used as controls. Total nitrogen (N) and ammonium N concentrations of the KGE were 1.47 and 1.46 g kg(-1), respectively. The bacteria tested were detected in both biogas effluents in the order of 2 to 3 log CFU g(-1), but there was little evidence that the biogas effluents increased these bacteria in the soil and the plant leaves. At the rate of 22 g N m(-2), yield, total N uptake, apparent N recovery rate, and leaf nitrate ion concentration at harvest of spinach and komatsuna in the KGE plot were mostly comparable to those in the NPK and CME plots. We conclude that the KGE is a quick-release N fertilizer comparable to chemical fertilizers and does not cause contamination of CG, E. coli, FS, or V. parahaemolyticus in the soil and spinach and komatsuna leaves.

Ectomycorrhizal (ECM) fungal mycelia are the main organs for nutrient uptake in many woody plants, and often connect seedlings to mature trees. While it is known that resources are shared among connected plants via common mycorrhizal networks (CMNs), the net effects of CMNs on seedling performance in the field are almost unknown. CMNs of individual ECM fungal species were produced in an early succession volcanic desert by transplanting current-year seedlings of Salix reinii with ECM mother trees that had been inoculated with one of 11 dominant ECM fungal species. Most seedlings were connected to individual CMNs without being infected by other ECM fungi. Although control seedlings showed poor growth under severe nutrient competition with larger nonmycorrhizal mother trees, nutrient acquisition and growth of seedlings connected to CMNs were improved with most fungal species. The positive effects of CMNs on seedling performance were significantly different among ECM fungal species; for example, the maximum difference in seedling nitrogen acquisition was 1 : 5.9. The net effects of individual CMNs in the field and interspecific variation among ECM fungal species are shown.

Sales and consumption of ready-to-eat aseptic steamed rice products have increased manyfold in Japan over the past 10 years. To determine the safety of steamed rice (water content 60%, pH 6.5) aseptically packaged under modified atmosphere, challenge studies were performed using a mixture of Clostridium botulinum proteolytic strains (five strains of type A and five strains of type B). Atmospheric conditions of 0 and 15% oxygen (with 5% CO2 and 5% N2 as the balance) were used. No neurotoxins were detected, and organoleptically acceptable conditions persisted for 24 weeks at 15% oxygen conditions. However, botulinum neurotoxin was found in one of three samples at 12 weeks and in one of two samples at 24 weeks at 0% oxygen and 30 degrees C. When samples were inoculated with C. botulinum with amylase (0% oxygen), neurotoxin and sample spoilage was detected after only 1 week of storage. Challenge studies using proteolytic strains of C. botulinum mixed with Bacillus subtilis (amylase formers) also were performed with atmosphere conditions of oxygen at 0, 5, 10, and 15% (with 5% CO2 and 5% N2 as the balance). Under 10 and 15% oxygen conditions, neurotoxin was not detected after 1 week of storage, but sample spoilage was detected after the same period. Under 0% oxygen conditions, neurotoxin was detected at 1 week, but the sample remained organoleptically acceptable even after 2 weeks of storage. Both neurotoxin and sample spoilage were detected at 1 week of storage under 5% oxygen conditions. Based on these results, cocontamination of amylase-producing Bacillus with C. botulinum would increase the risk of foodborne botulism when aseptic rice samples are packed under low-oxygen conditions (<5%). Therefore, to ensure the safety of these products, packing under atmospheric containing more than 10% oxygen is recommended.

We studied leaf area and nitrogen dynamics in the canopy of stands of an annual herb Xanthium canadense, grown at a high (HN)- and a low-nitorgen (LN) availability. Standing leaf area increased continuously through the vegetative growth period in the LN stand, or leveled off in the later stage in the HN stand. When scaled against standing leaf area, both production and loss rates of leaf area increased but with different patterns: the production rate was retarded, while the loss rate was accelerated, implying an upper limit of standing leaf area of the canopy. The rate of leaf-area production was higher in the HN than in the LN stand, which was caused by the higher rate of leaf production per standing leaf area as well as the greater standing leaf area in the HN stand. Although the rate of leaf-area loss was higher in the HN than in the LN stand, it was not significantly different between the two stands when compared at a common standing leaf area, suggesting involvement of light climate in determination of the leaf-loss rate. On the other hand, the rate of leaf-area loss was positively correlated with nitrogen demand for leaf area development across the two stands, suggesting that leaf loss was caused by retranslocation of nitrogen for construction of new leaves. A simple simulation model of leaf and nitrogen dynamics in the canopy showed that, at steady state, where the rate of leaf-area loss becomes equal to the production rate, the standing leaf area was still greater in the HN than in the LN stand. Similarly, when the uptake and loss of nitrogen are equilibrated, the standing nitrogen was greater in the HN than in the LN stand. These results suggest that leaf-area production is strongly controlled by nitrogen availability, while both nitrogen and light climate determine leaf-loss rates in the canopy.

This paper presents an evaluation of the process performance of a pilot-scale "fourth generation" downflow hanging sponge (DHS) post-treatment system combined with a UASB pretreatment unit treating municipal wastewater. After the successful operation of the second- and third-generation DHS reactors, the fourth-generation DHS reactor was developed to overcome a few shortcomings of its predecessors. This reactor was designed to further enhance the treatment efficiency and simplify the construction process in real scale, especially for the application in developing countries. Configuration of the reactor was modified to enhance the dissolution of air into the wastewater and to avert the possible clogging of the reactor especially during sudden washout from the UASB reactor. The whole system was operated at a total hydraulic retention time (HRT) of 8 h (UASB: 6 h and DHS: 2 h) for a period of over 600 days. The combined system was able to remove 96% of unfiltered BOD with only 9 mg/L remaining in the final effluent. Likewise, F. coli were removed by 3.45 log with the final count of 10(3) to 10(4) MPN/100 ml. Nutrient removal by the system was also satisfactory.

A mathematical model to estimate nitrogen (N) effluent from an upland field cultivated with barley, where under-drain pipes were installed 60 cm below the field surface, was developed and N effluents for several rainfall patterns during the cultivation period were analysed. The model is composed of the water drainage model and N cycle model. The water drainage model is made up of Sugawara's tank model in which the field is divided into two soil types, permeable soil and impermeable soil, and macro pore. The N cycle model can calculate the N reactions including nitrification, denitrification, mineralization, immobilization, urea hydrolysis and N transportation in the field. By using this model, N effluents caused by rainfall were analysed and characteristics of N effluent were clarified. The under drainages caused by heavy rains which occur around 60 days after fertilization contribute greatly to the amount of N effluent load during a cultivation period. Also, split application of fertilizer is not effective for the reduction of N effluents under any cultivation conditions, especially in winter crops. A large quantity of N effuses out of the field because the major portion of rainfall in a cultivation period occurs in the latter half of the period.

The aim of the study was to determine the concentrations of macronutrients and the mineral and trace element composition in maternal milk of Japanese women. We collected human milk samples from mothers living throughout Japan from December 1998 to September 1999, and defined as group A the 1197 samples among them that met the following conditions: breast milk of mothers who were under 40 years old, not in the habit of smoking and/or using vitamin supplements, and whose babies showed no symptoms of atopy and whose birth weights were 2.5 kg or more. We then analyzed their contents individually. We also analyzed the amino acid and free amino acid composition of the breast milk of pooled samples from various lactation stages. Large differences were found to exist among the contents of individual human milk samples. The mean contents of each component were as follows: energy, 66.3+/-13.3 kcal/100 mL; solid matter, 12.46+/-1.56 g/100 mL; ash, 0.19+/-0.06 g/100 mL; total nitrogen, 0.19+/-0.04 g/100 mL; lipids, 3.46+/-1.49 g/100 mL; carbohydrates, 7.58+/-0.77 g/100 mL; lactose, 6.44+/-0.49 g/100 mL; pH, 6.5+/-0.3; osmotic pressure, 299+/-14 mOsm/kg.H2O; chloride, 35.9+/-16.2 mg/100 mL; sodium, 13.5+/-8.7 mg/100 mL; magnesium, 2.7+/-0.9 mg/100 mL; phosphorus, 15.0+/-3.8 mg/100 mL; potassium, 47.0+/-12.1 mg/100 mL; calcium, 25.0+/-7.1 mg/100 mL; chromium, 5.9+/-4.7 microg/100 mL; manganese, 1.1+/-2.3 microg/100mL; iron, 119+/-251 microg/100 mL; copper, 35+/-21 microg/100 mL; zinc, 145+/-135 microg/100 mL; and selenium, 1.7+/-0.6 microg/100 mL. The content of each component varied greatly as the duration of lactation increased. In conclusion, it appears to be necessary to evaluate individual differences of human milk in order to perform valid research regarding infant formula.

This study was carried out to evaluate the quality of river water by analysis of land use in drainage basins and by estimating the N budgets. The drainage basins of Shibetsu River (Shibetsu area) and Bekkanbeushi River (Akkeshi area) in eastern Hokkaido, Japan were selected for a case study, and the evaluation of water quality was up-scaled to the regional level in Hokkaido by using the Arcview/GIS and statistical information. Water sampling was carried out in August 2001 and May 2002 in the Shibetsu and Akkeshi areas, respectively. The proportions of major land uses in drainage basins such as upland field, forest, urban area, wetland and wasteland for each sampling site were estimated by using topographic maps scaled at 1:25,000. The linear regression results showed that the correlation between NO3-N concentration and the proportion of upland in the drainage basins was highly and positively significant for both the Shibetsu area (r = 0.84, n = 57) and the Akkeshi area (r = 0.71, n = 73) at < 0.001 significance level. The regression coefficients or impact factors of river water quality were 0.015 and 0.0052 for the Shibetsu and Akkeshi areas, respectively. A comparison of these results with that of the previous study results in Hokkaido indicated that the impact factors were highest for intensive livestock farming areas (0.040), medium for mixed agriculture and livestock farming (0.020-0.030), and the lowest for grassland-based dairy cattle and horse farming areas (0.0052-0.015). The results of a simple regression analysis showed that the impact factors had a significant positive correlation with the cropland surplus N (r = 0.93, P < 0.01), chemical fertilizer N (r = 0.82, P < 0.05), and manure fertilizer N (r = 0.76, P < 0.05), which were estimated by using the N budget approach. Using the best-correlated regression model, impact factors for all cities, towns and villages of the Hokkaido region were estimated. The NO3-N concentrations for all major rivers in Hokkaido were predicted by multiplying the estimated impact factors by the proportion of uplands. The regression analysis indicated that the predicted NO3-N concentrations were significantly correlated (r = 0.62, P < 0.001, n = 203) with the measured NO3-N concentrations, reported previously. It can be concluded that estimating the proportions of upland fields in drainage basins, and calculating cropland surplus N enables us to predict river water quality with respect to NO3-N concentration.

In 1943, one of the authors (Miura) worked on the nutritional study of Japanese navy sailors. Fortunately the raw data of this study were conserved in secret in the private file of Miura. Recently, another author (Hashimoto) was seeking the data concerning nitrogen balance of Japanese people. Miura, recalling the remaining of the old data; he picked up the records of nitrogen metabolism of three sailors (A, B and C). Subject A, 34 years old, showed a net nitrogen balance of 77 g of protein per day; whereas subject B, 20 years old and subject C, 15 years old showed positive nitrogen balance. The authors remarked the relationship between unabsorbed nitrogen and nitrogen balance: The unabsorbed ratio of food nitrogen was the greatest in A (29%), but in B (22%) and C (19%) showed rather low ratio. Recent study using tracers, revealed that some amino acids, especially unabsorbed leucine in the intestine, might act as regulators of nitrogen balance in adults.

The present water quality conditions in the Seto Inland Sea of Japan are described along with an historical background before and after the measures taken to reduce eutrophication. The directive to reduce phosphorus discharge into this area was very effective, reducing the number of red tides from about 300 cases per year at their peak in 1976 to the recent level of about 100 cases per year, indicating the improvement of seawater quality. However, the hastiness of the measures taken to reduce phosphorous seems to have led to a depletion of dissolved inorganic phosphorus that is an essential nutrient for the growth of phytoplankton. Fishery production has also decreased with the reduction of phosphorus, showing a time lag, and the relationship between them shows a hysteresis-like pattern indicating that the condition of fishery production is currently in a critical stage of collapse. This implies that the phosphorus reduction could have lowered the phytoplankton primary production and also caused a detrimental effect on the fishery production. Noteworthy is the change in the phytoplankton species composition. The dominant species that form red tides have changed from non-harmful diatoms to harmful raphidophytes in the eutrophication process and then finally to harmful/toxic dinoflagellates in the oligotrophication process. This indicates that the measures to reduce phosphorus have caused a change in phytoplankton species composition, thereby altering the food web structure, suggesting that this might be the major cause of the reduction of fishery production. In conclusion, the Seto Inland Sea of Japan is apparently in an oligotrophic condition, and it could be said to be in the state of "cultural oligotrophication" caused by the hasty reduction of phosphorus loading. Dam construction, as another possible cause of the cultural oligotrophication, is also discussed, and evidence relating to the existence of numerous dams in this area is also shown. Stress is placed on the regulation of the level of nutrients and their elemental ratio in the seawater, because these factors are considered to be effective in preventing the generation of harmful algae and in sustaining the fishery production through preservation of the natural environment. A proposal is also described regarding how these factors could be controlled.

Experimental bioremediation of crude oil was conducted for approximately 3 months in the intertidal zone of the Sea of Japan, Hyogo Prefecture. Artificial mixtures of weathered Arabian light crude oil and sand taken from the experimental site were wrapped in polyester net envelopes. The envelopes were placed in drum-shaped acrylic vessels with perforated sides to facilitate seawater exchange. The vessels were laid in the intertidal area. Slow release nitrogen and phosphorus synthetic fertilizer granules were added to the oil-sand mixtures in three different amounts. Some oil-sand mixtures were unfertilized controls. The oil-sand mixtures were periodically sampled and changes in the composition of the residual oils were monitored. Oil samples were subjected to gas chromatography coupled with mass spectrometry for analysis of some representative semi-volatile aliphatic and aromatic compounds. All values for each analyte were normalized against that of hopane to evaluate the extent of oil biodegradation. Significant increases in the concentrations of both nitrogen and phosphorus were found in the fertilized sections in accordance with the amounts of added fertilizers. Although significant natural attenuation of oil was observed in the unfertilized sections, fertilization stimulated the degradation rate of the oil in the early stage of the experimental term. The extent of the oil biodegradation increased as the amount of added fertilizer increased. However, the final degradation efficiencies for each oil component in the fertilized sections were not significantly different from those in the unfertilized sections, and the degradation of each oil component had almost ceased after 6 weeks. We conclude that excessive amounts of macronutrients are required to accelerate oil biodegradation and that fertilization is only effective in the early stages.

The structure and function of Shisham (Dalbergia sissoo Roxb.) forests were investigated in relation to nutrient dynamics in 5- to 15-year-old stands growing in central Himalaya. Nutrient concentrations and storage in different layers of vegetation were in the order: tree > shrub > herb. Forest soil, litter and vegetation accounted for 80.1-91.9, 1.0-1.5 and 7.0-18.4%, respectively, of the total nutrients in the system. There were considerable reductions (trees 32.8-43.1; shrubs 26.2-32.4; and herbs 18-8-22-2%) in nutrient concentrations of leaves during senescence. Nutrient uptake by the vegetation as a whole and also by the different components, with and without adjustment for internal recycling, was investigated. Annual transfer of litter nutrients to the soil from vegetation was 74.8-108.4 kg ha(-1) year(-1) N, 56.8-4 kg ha(-1) year(-1) P and 38.7-46.9 kg ha(-1) year(-1) K. Turnover rate and time for different nutrients ranged between 56 and 66 % year(-1) and 1.5 and 1.8 years, respectively. The turnover rate of litter indicates that over 50% of nutrients in litter on the forest floor are released, which ultimately enhances the productivity of the forest stand. The nutrient use efficiency in Shisham forests ranged from 136 to 143 kg ha(-1) year(-1) for N, 1,441 to 1,570 kg ha(-1) year(-1) for P and 305 to 311 kg ha(-1) year(-1) for K. Compared with natural oak forest (265 kg ha(-1) year(-1) and an exotic eucalypt plantation (18 kg ha(-1) year(-1), a higher proportion of nutrients was retranslocated in Shisham forests, largely because of higher leaf tissue nutrient concentrations. This indicates a lower nutrient use efficiency of Shisham compared with eucalypt and oak. Compartment models for nutrient dynamics have been developed to represent the distribution of nutrients pools and net annual fluxes within the system.

The objective of this study is to find a possibility of complete treatment of garbage and resource recovery (production of methane from available utility of carbon resource in garbage) by biological treatment process. As the first step, a fact-finding survey of actual garbage discharged from the dormitory of the Ube National College of Technology (equivalent to 300 population) was carried out. Second, the combined biological anaerobic-aerobic treatment, i.e. combination of upflow anaerobic sludge blanket (UASB) process and aerobic membrane bioreactor (AMB) process, was applied to the garbage treatment. The applicability and efficiency of this system were investigated in this study. The survey results showed that the composition and quantity of garbage from a student dormitory changed slightly during a week due to the change of the menu, however, they remained almost unchanged during the entire experimental period. The experimental results showed high biodegradability of the garbage, and demonstrated its suitability for methane production. The soluble nitrogen removal was high: over 97%. No excess sludge was wasted from the system. A high treatment efficiency of simultaneous organic carbon and nitrogen was obtained. The possibility of complete treatment of garbage with this process has been positively demonstrated by this study.

The sufficiency of the dietary protein allowances recommended by the Ministry of Health and Welfare of Japan for various physical exercises has not been examined. We investigated the protein requirement for moderate-intensity physical exercise using nitrogen balance in the isoenergetic state. Seventeen healthy college students exercised for 10 days with an adaptation period of three days and non-exercise control period of 10 days. They were given a maintenance level of energy and 1.08 g/kg per day of high quality mixed proteins recommended by the Ministry of Health and Welfare of Japan throughout the experimental period. They performed treadmill running during the exercise period at about 65% of VO2max for 25, 41 or 48 min/day, which required 200, 300 or 400 kcal of extra energy, respectively. Although the exercise increased dermal nitrogen loss, urinary nitrogen excretion tended to decrease. Consequently, the exercise ranging from 200 to 400 kcal/day did not significantly alter the nitrogen balance. Thiobarbituric acid-reactive substances and glutathione disulfide were not increased after exercise, suggesting that the moderate exercise, recommended for health promotion by the Ministry of Health and Welfare of Japan, does not cause oxidative damage to healthy subjects. These findings demonstrate that a protein intake of 1.08 g/kg per day is enough for moderate exercise.

To investigate genetic polymorphism of N-acetylation in Japanese population we measured isoniazid (INH) and acetylisoniazid (AcINH) in the urine of 340 unrelated Japanese subjects. For the detection of polymorphism, we used probit analysis, logit analysis, and arcsine transformation of the logarithm of molar acetylation ratio (log (INH/AcINH)). The observed antimodes were -0.85 and -0.325 for the probit, 4.899 and 7.154 for the logit, 43.85 and 71.57 for the arcsine analysis plot. Probit and arcsine analysis were sensitive for the separation of intermediate from slow acetylators, while logit analysis was sensitive for the separation of rapid from intermediate acetylators. Clinically it is important to know to which genetic group the patient belongs because we may have to increase the dosage for the rapid acetylators to achieve the desired therapeutic effect, while we may have to reduce the dosage for slow acetylators in whom the incidence of side effect is high.

BOD (Biochemical Oxygen Demand) is utilized widely as an index of water quality. However, nitrification is a cause of significant errors in measuring BOD, particularly when a large population of nitrifying organisms are existing in water such as effluents from biological treatment plants. In this case, the use of a nitrification inhibitor is recommended, but it is not used commonly. In order to investigate the amount of nitrogenous oxygen demand (NOD) which is caused by nitrification, BOD with and without inhibitor was measured as samples in the Kitakyushu area. About 70 percent of BOD from the effluents of biological treatment plants was NOD. In the case of influent of biological treatment plant and river waters, 0-39 percent of NOD was contained in BOD.

Obligatory urinary and fecal N losses in young Japanese women were evaluated and the effect of energy intake on the utilization of rice and egg mixed protein was investigated in subjects receiving a low protein diet. Seven female students were given a protein-free diet for ten days. Feces and 24-h urine samples were collected throughout the period and their nitrogen contents were analyzed. Basal metabolism was measured and the lean body mass was calculated from urinary creatinine excretion by the equation of Forbes. The mean weight loss of the 7 subjects during the ten days was 1.5 kg, the mean obligatory urinary N loss was 32.3 +/- 5.9 mg/kg body weight, 1.67 +/- 0.45 mg/kcal basal metabolism, or 1.68 +/- 0.32 g/g creatinine, and the mean obligatory fecal N loss was 10.1 +/- 2.0 mg/kg body weight, or 0.51 +/- 0.08 mg/kcal basal metabolism. In a second experiment, 17 female subjects were divided into three groups with an energy intake of about 35, 42, 46 kcal/kg BW, respectively. During the 7-day experimental period, they were given a low protein diet containing 250 g of rice and 125 g of whole egg. The nitrogen contents of the 24-h urine samples and feces collected were analyzed and the nitrogen balance was calculated as the difference between N intake and urinary and fecal N excretion. Subjects with an energy intake of 35 kcal/kg BW had a negative N balance, while subjects with an intake of 46 kcal/kg BW achieved an apparent positive N balance. The NPU values of rice and egg mixed protein with energy intake of 35, 42, 46 kcal/kg BW were calculated from the N balance data and the values for obligatory urinary and fecal N losses in the first experiment as 25, 37 and 54, respectively.

Eighteen refractory obese patients received a formula diet of 240 kcal per day using Modifast (VLCD 240) for 1-2 months of patient treatment. Another 22 obese patients were treated by Optifast 70, which supplies 420 kcal per day (VLCD 420), for 1-2 months on an inpatient or outpatient basis. The weight losses attained through these VLCD regimens during the first month of treatment were compared. In addition, the metabolic effects of each VLCD regimen were investigated in 21 inpatients, of whom 11 received the VLCD 240 and 10 received the VLCD 420. The nitrogen balance, levels of serum rapid turnover proteins and serum total ketone bodies concentration of each individual treated by the two different VLCD regimens were also investigated. The VLCD 420 showed an identical weight reduction effect (7.6 +/- 2.4 kg/month) to that attained by the VLCD 240 (8.9 +/- 3.1 kg/month). The nitrogen balance turned out to be negative in each VLCD regimen for the first week of treatment. The VLCD 240 negative nitrogen balance continued to the end of the fourth week, but the VLCD 420 negative nitrogen balance recovered to a state of equilibrium around the third or fourth weeks. The serum prealbumin and retinol-binding protein levels decreased rapidly after the initiation of each VLCD regimen; however, a prominent decrease of the serum retinol-binding protein level was observed with the VLCD 240. A positive correlation between the total nitrogen loss and the decrease of serum prealbumin concentration was observed during the first month of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)