Impact of climate change and oligotrophication on quality and quantity of lake primary production: A case study in Lake Biwa.

Global climate change and anthropogenic oligotrophication are expected to reshape the dynamics of primary production (PP) in aquatic ecosystems; however, few studies have explored their long-term effects. In theory, the PP of phytoplankton in Lake Biwa may decline over decades due to warming, heightened stratification, and anthropogenic oligotrophication. Furthermore, the PP of large phytoplankton, which are inedible to zooplankton, along with biomass-specific productivity (PBc), could decrease. In this study, data from 1976 to 2021 and active fluorometry measurements taken in 2020 and 2021 were evaluated. Quantitatively, the temporal dynamics of mean seasonal PP during 1971-2021 were assessed according to the carbon fixation rate to investigate relationships among environmental factors. Qualitatively, phytoplankton biomass, PP, and PBc were measured in two size fractions [edible (S) or inedible (L) for zooplankton] in 2020 and 2021, and the L:S balance for these three measures was compared between 1992 (low-temperature/high-nutrient conditions) and 2020-2021 (high-temperature/low-nutrient conditions) to assess seasonal dynamics. The results indicated that climate change and anthropogenic oligotrophication over the past 30 years have diminished Lake Biwa's PP since the 1990s, impacting the phenology of PP dynamics. However, the L:S balance in PP and PBc has exhibited minimal change between the data from 1992 and the 2020-2021 period. These findings suggest that, although climate change and oligotrophication may reduce overall PP, they may not markedly alter the inedible/edible phytoplankton balance in terms of PP and PBc. Instead, as total PP declines, the production of small edible phytoplankton may decrease proportionally, potentially affecting trophic transfer efficiency and material cycling in Lake Biwa.

Identifying the true drivers of abrupt changes in ecosystem state with a focus on time lags: Extreme precipitation can determine water quality in shallow lakes.

A better understanding of abrupt ecosystem changes is needed to improve prediction of future ecosystem states under climate change. Chronological analysis based on long-term monitoring data is an effective way to estimate the frequency and magnitude of abrupt ecosystem changes. In this study, we used abrupt-change detection to differentiate changes of algal community composition in two Japanese lakes and to identify the causes of long-term ecological transitions. Additionally, we focused on finding statistically significant relationships between abrupt changes to aid with factor analysis. To estimate the strengths of driver-response relationships underlying abrupt algal transitions, the timing of the algal transitions was compared to that of abrupt changes in climate and basin characteristics to identify any synchronicities between them. The timing of abrupt algal changes in the two study lakes corresponded most closely to that of heavy runoff events during the past 30-40 years. This strongly suggests that changes in the frequency of extreme events (e.g., heavy rain, prolonged drought) have a greater effect on lake chemistry and community composition than do shifts in the means of climate and basin factors. Our analysis of synchronicity (with a focus on time lags) could provide an easy method to identify better adaptative strategies for future climate change.

Measuring Photophysiology of Attached Stage of Colacium sp. by a Cuvette-Type Fast Repetition Rate Fluorometer.

Fast repetition rate fluorometer (FRRf) is a beneficial method for measuring photosystem II (PSII) photophysiology and primary productivity. Although FRRf can measure PSII absorption cross-section (σPSII), maximum photochemical efficiency (Fv/Fm), effective photochemical efficiency (Fq'/Fm'), and non-photochemical quenching (NPQNSV) for various eukaryotic algae and cyanobacteria, almost all FRRf studies to date have focused on phytoplankton. Here, the protocol describes how to measure PSII photophysiology of an epizoic alga Colacium sp. Ehrenberg 1834 (Euglenophyta), in its attached stage (attached to zooplankton), using cuvette-type FRRf. First, we estimated the effects of substrate zooplankton (Scapholeberis mucronata O.F. Müller 1776, Cladocera, Daphniidae) on baseline fluorescence and σPSII, Fv/Fm, Fq'/Fm', and NPQNSV of planktonic Colacium sp. To validate this methodology, we recorded photophysiology measurements of attached Colacium sp. on S. mucronata and compared these results with its planktonic stage. Representative results showed how the protocol could determine the effects of calcium (Ca) and manganese (Mn) on Colacium sp. photophysiology and identify the various effects of Mn enrichment between attached and planktonic stages. Finally, we discuss the adaptability of this protocol to other periphytic algae.

Development of photosynthetic carbon fixation model using multi-excitation wavelength fast repetition rate fluorometry in Lake Biwa.

Direct measurements of gross primary productivity (GPP) in the water column are essential, but can be spatially and temporally restrictive. Fast repetition rate fluorometry (FRRf) is a bio-optical technique based on chlorophyll a (Chl-a) fluorescence that can estimate the electron transport rate (ETRPSII) at photosystem II (PSII) of phytoplankton in real time. However, the derivation of phytoplankton GPP in carbon units from ETRPSII remains challenging because the electron requirement for carbon fixation (Фe,C), which is mechanistically 4 mol e- mol C-1 or above, can vary depending on multiple factors. In addition, FRRf studies are limited in freshwater lakes where phosphorus limitation and cyanobacterial blooms are common. The goal of the present study is to construct a robust Фe,C model for freshwater ecosystems using simultaneous measurements of ETRPSII by FRRf with multi-excitation wavelengths coupled with a traditional carbon fixation rate by the 13C method. The study was conducted in oligotrophic and mesotrophic parts of Lake Biwa from July 2018 to May 2019. The combination of excitation light at 444, 512 and 633 nm correctly estimated ETRPSII of cyanobacteria. The apparent range of Фe,C in the phytoplankton community was 1.1-31.0 mol e- mol C-1 during the study period. A generalised linear model showed that the best fit including 12 physicochemical and biological factors explained 67% of the variance in Фe,C. Among all factors, water temperature was the most significant, while photosynthetically active radiation intensity was not. This study quantifies the in situ FRRf method in a freshwater ecosystem, discusses core issues in the methodology to calculate Фe,C, and assesses the applicability of the method for lake GPP prediction.

Primary production estimated for large lakes and reservoirs in the Mekong River Basin.

Understanding the proximate factors and mechanisms driving primary production in manmade reservoirs is crucial because such production can translate into added fish yields that provide people with food and livelihoods. Furthermore, reservoir fish production could potentially compensate for the loss of fish yields due to habitat fragmentation and alterations caused by damming and impoundment. We monitored primary production, identified environmental factors responsible for its variability, and examined the relationship between primary production and fish production in nine large water bodies of the Lower Mekong Basin for 2 years. The estimated primary production ranged from 40 to 302 g C/m2/y and was generally greater in the wet season than in the dry season. Linear mixed-effects modelling identified the concentration of dissolved inorganic carbon as a significant fixed-effect variable regulating primary production, after variability due to random and fixed effects of water body and seasonality, respectively, were taken into account. Fish yields marginally increased with increasing primary production across the water bodies, with the estimated energy transfer efficiency ranging from 0.004 to 0.009. Dissolved inorganic carbon was partly determined by the lithological composition of the water body catchment, suggesting that the geographic locations of proposed dams determine the magnitude of primary production and hence future fish production.

Magnetic Resonance Imaging as a Novel Method for Elucidating Sediment Burrow Structures and Functions.

Burrow structures produced by various benthic animals in sediments are important components of aquatic ecosystems, allowing the circulation of interstitial water via ingress of fresh bottom water into the burrows upon feeding and intraburrow migration. Although X-ray computed tomography has been used to visualize burrow structures, it could not reveal the structures in the soft mud in Lake Kasumigaura, where evaluation of the water-circulation effect of burrows is an important issue. Here, we describe the first attempt to use magnetic resonance (MR) imaging (MRI) to visualize intact burrow structures in the soft mud sediment cores collected from a eutrophic lake. Our MRI application clarified the dynamic distribution of burrows inhabited by chironomids in the soft mud that previous studies could not visualize. By examining the relationships between the degree of chloride ion depletion in deeper layers and the burrow density calculated from the MR images, we were able to consistently explain the water-circulation effect of burrows, suggesting the higher reliability of burrow density calculated from MR images. In addition, we were able to evaluate the activity of burrows, which is difficult to achieve in sediment core experiments. We observed a smaller water-circulation effect of burrows on ammonium ions than on chloride ions, suggesting the enhancement of ammonium production or release in burrow-rich sediments.

Incorporation characteristics of exogenous 15N-labeled thymidine, deoxyadenosine, deoxyguanosine and deoxycytidine into bacterial DNA.

Bacterial production has been often estimated from DNA synthesis rates by using tritium-labeled thymidine. Some bacteria species cannot incorporate extracellular thymidine into their DNA, suggesting their biomass production might be overlooked when using the conventional method. In the present study, to evaluate appropriateness of deoxyribonucleosides for evaluating bacterial production of natural bacterial communities from the viewpoint of DNA synthesis, incorporation rates of four deoxyribonucleosides (thymidine, deoxyadenosine, deoxyguanosine and deoxycytidine) labeled by nitrogen stable isotope (15N) into bacterial DNA were examined in both ocean (Sagami Bay) and freshwater (Lake Kasumigaura) ecosystems in July 2015 and January 2016. In most stations in Sagami Bay and Lake Kasumigaura, we found that incorporation rates of deoxyguanosine were the highest among those of the four deoxyribonucleosides, and the incorporation rate of deoxyguanosine was approximately 2.5 times higher than that of thymidine. Whereas, incorporation rates of deoxyadenosine and deoxycytidine were 0.9 and 0.2 times higher than that of thymidine. These results clearly suggest that the numbers of bacterial species which can incorporate exogenous deoxyguanosine into their DNA are relatively greater as compared to the other deoxyribonucleosides, and measurement of bacterial production using deoxyguanosine more likely reflects larger numbers of bacterial species productions.

Characterization of dissolved organic matter in wastewater during aerobic, anaerobic, and anoxic treatment processes by molecular size and fluorescence analyses.

Changes in the characteristics of dissolved organic matter (DOM: the dissolved fraction of natural organic matter) during a series of wastewater treatment plant (WWTP) processes were investigated by using a combination of molecular size analysis and excitation emission matrix (EEM) spectroscopy coupled with parallel factor analysis. The characteristics of DOM were compared following aerobic, anoxic, and anaerobic treatments. Three peaks at about 100,000 Da (high-molecular-size DOM, Peak 1) and about 900-1,100 Da (intermediate-molecular-size DOM, Peak 2; low-molecular-size DOM, Peak 3 as the shoulder of Peak 2) were observed in the distribution of total organic carbon molecular sizes in the influent of the WWTPs. In this study, five fluorescent components (C1 to C5) were identified in the EEM spectra. Molecular size analysis and molecular size fractionation revealed that the C3 (humic-like) and C5 (specific to sewage) fluorophores had intermediate or low molecular sizes. Comparison of the changes of the concentrations of dissolved organic carbon in each reaction tank and investigation of the removal selectivity of each treatment (aerobic, anaerobic, and anoxic) suggested that the heterogenous compounds present in DOM of the influent were homogenized into intermediate-molecular-size DOM with high hydrophobicity and aromaticity, or into C4 fluorophores (DOM-X), during anaerobic or anoxic treatment. DOM-X was able to be transformed or removed by aerobic treatment. The results suggested that introduction of aerobic treatment at the appropriate stage of wastewater treatment or inclusion of physical or chemical treatment should be an effective way to optimize DOM removal.

Vertical variation of bulk and metabolically active prokaryotic community in sediment of a hypereutrophic freshwater lake.

This study was conducted to acquire novel insight into differences between bulk (16S rDNA) and metabolically active (16S rRNA) prokaryotic communities in the sediment of a hypereutrophic lake (Japan). In the bulk communities, the class Deltaproteobacteria and the order Methanomicrobiales were dominant among bacteria and methanogens. In the metabolically active communities, the class Alphaproteobacteria and the order Methanomicrobiales and the family Methanosaetaceae were frequently found among bacteria and methanogens. Unlike the bulk communities of prokaryotes, the composition of the metabolically active communities varied remarkably vertically, and their diversities greatly decreased in the lower 20 cm of sediment. The metabolically active prokaryotic community in the sediment core was divided into three sections based on their similarity: 0-6 cm (section 1), 9-18 cm (section 2), and 21-42 cm (section 3). This sectional distribution was consistent with the vertical pattern of the sedimentary stable carbon and nitrogen isotope ratios and oxidation-reduction potential in the porewater. These results suggest that vertical disturbance of the sediment may influence the communities and functions of metabolically active prokaryotes in freshwater lake sediments. Overall, our results indicate that rRNA analysis may be more effective than rDNA analysis for evaluation of relationships between actual microbial processes and material cycling in lake sediments.

Coniferous coverage as well as catchment steepness influences local stream nitrate concentrations within a nitrogen-saturated forest in central Japan.

High concentrations of nitrate have been detected in streams flowing from nitrogen-saturated forests; however, the spatial variations of nitrate leaching within those forests and its causes remain poorly explored. The aim of this study is to evaluate the influences of catchment topography and coniferous coverage on stream nitrate concentrations in a nitrogen-saturated forest. We measured nitrate concentrations in the baseflow of headwater streams at 40 montane forest catchments on Mount Tsukuba in central Japan, at three-month intervals for 1 year, and investigated their relationship with catchment topography and with coniferous coverage. Although stream nitrate concentrations varied from 0.5 to 3.0 mgN L-1, those in 31 catchments consistently exceeded 1 mgN L-1, indicating that this forest had experienced nitrogen saturation. A classification and regression tree analysis with multiple environmental factors showed that the mean slope gradient and coniferous coverage were the best and second best, respectively, at explaining inter-catchment variance of stream nitrate concentrations. This analysis suggested that the catchments with steep topography and high coniferous coverage tend to have high nitrate concentrations. Moreover, in the three-year observation period for five adjacent catchments, the two catchments with relatively higher coniferous coverage consistently had higher stream nitrate concentrations. Thus, the spatial variations in stream nitrate concentrations were primarily regulated by catchment steepness and, to a lesser extent, coniferous coverage in this nitrogen-saturated forest. Our results suggest that a decrease in coniferous coverage could potentially contribute to a reduction in nitrate leaching from this nitrogen-saturated forest, and consequently reduce the risk of nitrogen overload for the downstream ecosystems. This information will allow land managers and researchers to develop improved management plans for this and similar forests in Japan and elsewhere.

The dynamics of pico-sized and bloom-forming cyanobacteria in large water bodies in the Mekong River Basin.

In the face of plans for increased construction of dams and reservoirs in the Mekong River Basin, it is critically important to better understand the primary-producer community of phytoplankton, especially the warm-water cyanobacteria. This is because these algae can serve as the primary source of carbon for higher trophic levels, including fishes, but can also form harmful blooms, threatening local fisheries and environmental and human health. We monitored the dynamics of three cyanobacteria-Synechococcus spp., Microcystis aeruginosa, and Dolichospermum spp.-for two years in nine large lakes and reservoirs in the Mekong River Basin. The densities of these algae were largely system-specific such that their abundance was uniquely determined within individual water bodies. However, after accounting for the system-specific effect, we found that cell densities of Synechococcus spp., M. aeruginosa, and Dolichospermum spp. varied in response to changes in photosynthetically active radiation (PAR), total nitrogen, and water level, respectively. Because both PAR and water level tend to fluctuate concordantly over a wide geographic area, Synechococcus spp., and to a lesser extent Dolichospermum spp., varied synchronously among the water bodies. Sustaining the production of pico-sized primary producers while preventing harmful algal blooms will be a key management goal for the proposed reservoirs in the Mekong Basin.

Improvement of COD removal by controlling the substrate degradability during the anaerobic digestion of recalcitrant wastewater.

The recalcitrant landfill leachate was anaerobically digested at various mixing ratios with labile synthetic wastewater to evaluate the degradation properties of recalcitrant wastewater. The proportion of leachate to the digestion system was increased in three equal steps, starting from 0% to 100%, and later decreased back to 0% with the same steps. The chemical oxygen demand (COD) for organic carbon and other components were calculated by analyzing the COD and dissolved organic carbon (DOC), and the removal efficiencies of COD carbon and COD others were evaluated separately. The degradation properties of COD carbon and COD others shifted owing to changing of substrate degradability, and the removal efficiencies of COD carbon and COD others were improved after supplying 100% recalcitrant wastewater. The UV absorptive property and total organic carbon (TOC) of each molecular size using high performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) with UVA and TOC detectors were also investigated, and the degradability of different molecular sizes was determined. Although the SEC system detected extracellular polymeric substances (EPS), which are produced by microbes in stressful environments, during early stages of the experiment, EPS were not detected after feeding 100% recalcitrant wastewater. These results suggest that the microbes had acclimatized to the recalcitrant wastewater degradation. The high removal rates of both COD carbon and COD others were sustained when the proportion of labile wastewater in the substrate was 33%, indicating that the effective removal of recalcitrant COD might be controlled by changing the substrate's degradability.

Dynamics of particulate phosphorus in a shallow eutrophic lake.

We tested the hypothesis that in shallow, eutrophic Lake Kasumigaura, the concentration of particulate phosphorus (PP) is controlled by biogenic P (P in living or dead phytoplankton and bacterial cells), rather than by resuspension of inorganic P in sediment. Increases in wind velocity and turbidity were associated with bottom shear stress exceeding the critical value for the lake (τc=0.15Nm(-2)); this increased turbidity was due to sediment resuspension. However, concentrations of PP; HCl-extractable, reactive P in PP (P-rP); and HCl-extractable, non-reactive P in PP (P-nrP) were not correlated with wind velocity (PP vs. wind velocity: r=0.40, p>0.05). Rather, the P-nrP concentration accounted for approximately 79% of PP, and the concentrations of PP, P-rP, and P-nrP were correlated with the particulate organic carbon (POC) concentration (POC vs. PP: r=0.90, p<0.01; POC vs. P-rP: r=0.82, p<0.01; POC vs. P-nrP: r=0.86, p<0.01). In our (31)P nuclear magnetic resonance spectroscopy results, mononucleotides accounted for the largest proportion among the detected P compound classes. In addition, concentrations of mononucleotides, orthophosphate, and pyrophosphate were significantly higher in samples with high POC concentrations, whereas the DNA-P concentration was not. These results suggest that biogenic P affects PP concentrations more strongly than does sediment resuspension, and the production of biogenic P creates a pool of mononucleotides, a class of easily degradable P, even in shallow, eutrophic Lake Kasumigaura.

Unexpected Diversity of pepA Genes Encoding Leucine Aminopeptidases in Sediments from a Freshwater Lake.

We herein designed novel PCR primers for universal detection of the pepA gene, which encodes the representative leucine aminopeptidase gene, and investigated the genetic characteristics and diversity of pepA genes in sediments of hypereutrophic Lake Kasumigaura, Japan. Most of the amino acid sequences deduced from the obtained clones (369 out of 370) were related to PepA-like protein sequences in the M17 family of proteins. The developed primers broadly detected pepA-like clones associated with diverse bacterial phyla-Alpha-, Beta-, Gamma-, and Deltaproteobacteria, Acidobacteria, Actinobacteria, Aquificae, Chlamydiae, Chloroflexi, Cyanobacteria, Firmicutes, Nitrospirae, Planctomycetes, and Spirochetes as well as the archaeal phylum Thaumarchaeota, indicating that prokaryotes in aquatic environments possessing leucine aminopeptidase are more diverse than previously reported. Moreover, prokaryotes related to the obtained pepA-like clones appeared to be r- and K-strategists, which was in contrast to our previous findings showing that the neutral metalloprotease gene clones obtained were related to the r-strategist genus Bacillus. Our results suggest that an unprecedented diversity of prokaryotes with a combination of different proteases participate in sedimentary proteolysis.

Production of renewable hexanols from mechanocatalytically depolymerized cellulose by using Ir-ReOx /SiO2 catalyst.

Hexanols were produced in high yield by conversion of cellulose over Ir-ReOx /SiO2 (molar ratio Re/Ir=2) catalyst in biphasic reaction system (n-decane+H2 O). The cellulose was depolymerized by mechanocatalysis with the aid of H2 SO4 . The influence of solvent amount, reaction temperature and hydrogen pressure was systematically investigated and the highest yield of hexanols reached 60 % under the conditions of n-decane/water ∼2 (v/v), 413 K, 10 MPa H2 for 24 h. Mechanocatalytic depolymerization of cellulose with the aid of H2 SO4 or HCl and the use of sufficient n-decane were very crucial for the production of hexanols. H2 SO4 not only catalyzed cellulose to water-soluble oligosaccharides but also promoted the hydrogenolysis activity of Ir-ReOx /SiO2 catalyst. The role of n-decane was to extract hexanols and to suppress over-hydrogenolysis of hexanols to n-hexane.

Linking temporal changes in bacterial community structures with the detection and phylogenetic analysis of neutral metalloprotease genes in the sediments of a hypereutrophic lake.

We investigated spatial and temporal variations in bacterial community structures as well as the presence of three functional proteolytic enzyme genes in the sediments of a hypereutrophic freshwater lake in order to acquire an insight into dynamic links between bacterial community structures and proteolytic functions. Bacterial communities determined from 16S rRNA gene clone libraries markedly changed bimonthly, rather than vertically in the sediment cores. The phylum Firmicutes dominated in the 4-6 cm deep sediment layer sample after August in 2007, and this correlated with increases in interstitial ammonium concentrations (p < 0.01). The Firmicutes clones were mostly composed of the genus Bacillus. npr genes encoding neutral metalloprotease, an extracellular protease gene, were detected after the phylum Firmicutes became dominant. The deduced Npr protein sequences from the retrieved npr genes also showed that most of the Npr sequences used in this study were closely related to those of the genus Bacillus, with similarities ranging from 61% to 100%. Synchronous temporal occurrences of the 16S rRNA gene and Npr sequences, both from the genus Bacillus, were positively associated with increases in interstitial ammonium concentrations, which may imply that proteolysis by Npr from the genus Bacillus may contribute to the marked increases observed in ammonium concentrations in the sediments. Our results suggest that sedimentary bacteria may play an important role in the biogeochemical nitrogen cycle of freshwater lakes.

Bacterial contribution to dissolved organic matter in eutrophic Lake Kasumigaura, Japan.

Incubation experiments using filtered waters from Lake Kasumigaura were conducted to examine bacterial contribution to a dissolved organic carbon (DOC) pool. Bacterial abundance, bacterial production, concentrations of DOC, total dissolved amino acids (TDAA), and total dissolved neutral sugars (TDNS) were monitored during the experiments. Bacterial production during the first few days was very high (20 to 35 µg C liter(-1) day(-1)), accounting for 40 to 70% of primary production. The total bacterial production accounted for 34 to 55% of the DOC loss during the experiment, indicating high bacterial activities in Lake Kasumigaura. The DOC degradation was only 12 to 15%, whereas the degradation of TDAA and TDNS ranged from 30 to 50%, suggesting the preferential usage of TDAA and TDNS. The contribution of bacterially derived carbon to a DOC pool in Lake Kasumigaura was estimated using d-amino acids as bacterial biomarkers and accounted for 30 to 50% of the lake DOC. These values were much higher than those estimated for the open ocean (20 to 30%). The ratio of bacterially derived carbon to bulk carbon increased slightly with time, suggesting that the bacterially derived carbon is more resistant to microbial degradation than bulk carbon. This is the first study to estimate the bacterial contribution to a DOC pool in freshwater environments. These results indicate that bacteria play even more important roles in carbon cycles in freshwater environments than in open oceans and also suggests that recent increases in recalcitrant DOC in various lakes could be attributed to bacterially derived carbon. The potential differences in bacterial contributions to dissolved organic matter (DOM) between freshwater and marine environments are discussed.

Biogenic phosphorus compounds in sediment and suspended particles in a shallow eutrophic lake: a ³¹P-nuclear magnetic resonance (³¹P NMR) study.

Differences in biogenic phosphorus (P) compounds between sediment and suspended particles in aquatic environments are important for understanding the mechanisms of internal P loading, but these differences are still unknown. We used solution-state (31)P-nuclear magnetic resonance spectroscopy ((31)P NMR) with NaOH-ethylenediaminetetraacetic extraction to detect the multiple P compounds in suspended particles and sediment in the eutrophic Lake Kasumigaura, including orthophosphate monoesters, orthophosphate diesters, pyrophosphate, and polyphosphate. We tested the hypothesis that there is a significant difference between these groups in suspended particles and sediment. Biogenic P other than orthophosphate was found in significantly higher proportions in suspended particles (74.3% of total P) than in sediment (25.6%). Orthophosphate monoesters were comparatively more abundant in suspended particles, as indicated by the ratio of orthophosphate diesters to monoesters (average, 0.31 for suspended particles; 1.05 for sediment). The compounds identified as orthophosphate monoesters by (31)P NMR spectroscopy originated mainly from phospholipids (α-glycerophosphate and ß-glycerophosphate) and ribonucleic acid (RNA-P), whereas the orthophosphate diesters included mostly DNA (DNA-P). These results suggest that the dynamics of orthophosphate diesters, the production of DNA-P, or the degradation of phospholipids, play an important role in P cycling in Lake Kasumigaura.

Ecological niche separation in the Polynucleobacter subclusters linked to quality of dissolved organic matter: a demonstration using a high sensitivity cultivation-based approach.

The free-living, cosmopolitan, freshwater betaproteobacterial bacterioplankton genus Polynucleobacter was detected in different years in 11 lakes of varying types and a river using the size-exclusion assay method (SEAM). Of the 350 strains isolated, 228 (65.1%) were affiliated with the Polynucleobacter subclusters PnecC (30.0%) and PnecD (35.1%). Significant positive correlations between fluorescence in situ hybridization and SEAM data were observed in the relative abundance of PnecC and PnecD bacteria to Polynucleobacter communities (PnecC + PnecD). Isolates were mainly PnecC bacteria in the samples with a high specific UV absorbance at 254 nm (SUVA(254) ), and a low total hydrolysable neutral carbohydrate and amino acid (THneutralCH + THAA) content of the dissolved organic matter (DOM) fraction, which is known to be correlated with a high humic content. In contrast, the PnecD bacteria were abundant in samples with high chlorophyll a and/or THneutralCH + THAA concentrations, indicative of primary productivity. With few exceptions, differences in the relative abundance of PnecC and PnecD in each sample, determined using a high-sensitivity cultivation-based approach, were due to DOM quality. These results suggest that the major DOM component in the field, which is allochthonously or autochthonously derived, is a key factor for ecological niche separation between PnecC and PnecD subclusters.

Fast and precise method for HPLC-size exclusion chromatography with UV and TOC (NDIR) detection: importance of multiple detectors to evaluate the characteristics of dissolved organic matter.

A new type of high-performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) system with ultraviolet (UV) absorbance detection and non-dispersive infrared (NDIR) detection of total organic carbon is described. The introduction of an online degassing tube and a low-volume HPLC column helped to reduce the analytical time and increase the sensitivity of the SEC system. This study is the first in which linear calibration curves (R(2)>0.99) were obtained for both UV absorbance and NDIR data for polystyrene sulfonate standards, which are the most suitable standards for molecular size analysis of aquatic humic substances as well as dissolved organic matter (DOM). Using the calibration curves, the molecular size distribution of DOM in water collected from Lake Kasumigaura and in pore water from lake sediments was estimated. Most of the DOM had a molecular weight less than 4000 Daltons (Da), and the amount of low-molecular-weight DOM (∼ 2000 Da) with low UV absorbance increased with depth in the sediment pore water. This result shows the importance of combining quantitative analysis by NDIR detection with qualitative analysis by UV detection to determine the chemical and physical properties of DOM. The possible sources and reactivity of DOM in Lake Kasumigaura and its sediment pore water are also discussed.