Temporal and spatial variations in the composition of freshwater photosynthetic picoeukaryotes revealed by MiSeq sequencing from flow cytometry sorted samples.

The diversity and composition of photosynthetic picoeukaryotes (PPEs) in two large shallow lakes in China (Lake Taihu and Lake Chaohu) were investigated from flow cytometry sorted samples using Miseq high-throughput sequencing. We collected 65 samples covering different regions of the two lakes over four seasons to unveil spatial and temporal patterns of PPEs community composition. The use of flow cytometry sorting largely improved the efficiency of detecting PPEs sequences and over 70% of the retrieved reads belonged to PPEs. Chlorophyta and Bacillariophyta dominated PPEs in most of the samples. A distinct but complex seasonality of PPEs composition emerged at the OTUs level. NGS-based Miseq sequencing facilitates an in-depth view of numerous rare OTUs. Nearly 80% of the PPEs OTUs were rare and lots of them were detected only in one season, whereas most of the abundant OTUs were frequently detected in all seasons but only changed in relative abundances. Besides, a close relative of the marine PPEs species Ostreococcus sp. (OTU_1144, 99% identity) was discovered in freshwater systems for the first time and was abundant especially in winter. The diversity and community composition of PPEs were more dependent on season rather than sampling sites. Temperature, phycocyanin and NO3 N concentrations in Lake Taihu explained the PPE composition variations, whereas in Lake Chaohu TN/TP ratios, temperature, pH and nephelometric turbidity units (NTU) seemed to be the most important factors. In addition, a great number of OTUs belong to nonpigmented picoeukaryotes, especially Chytridiomycota, Perkinsozoa, Ciliophora and Cercozoa, which are known to include algae parasites as well as predators. The results of mantel test also showed that the community of photosynthetic and nonpigmented picoeukaryotes were significantly correlated in both lakes.

Hydrological regulation drives regime shifts: evidence from paleolimnology and ecosystem modeling of a large shallow Chinese lake.

Quantitative evidence of sudden shifts in ecological structure and function in large shallow lakes is rare, even though they provide essential benefits to society. Such 'regime shifts' can be driven by human activities which degrade ecological stability including water level control (WLC) and nutrient loading. Interactions between WLC and nutrient loading on the long-term dynamics of shallow lake ecosystems are, however, often overlooked and largely underestimated, which has hampered the effectiveness of lake management. Here, we focus on a large shallow lake (Lake Chaohu) located in one of the most densely populated areas in China, the lower Yangtze River floodplain, which has undergone both WLC and increasing nutrient loading over the last several decades. We applied a novel methodology that combines consistent evidence from both paleolimnological records and ecosystem modeling to overcome the hurdle of data insufficiency and to unravel the drivers and underlying mechanisms in ecosystem dynamics. We identified the occurrence of two regime shifts: one in 1963, characterized by the abrupt disappearance of submerged vegetation, and another around 1980, with strong algal blooms being observed thereafter. Using model scenarios, we further disentangled the roles of WLC and nutrient loading, showing that the 1963 shift was predominantly triggered by WLC, whereas the shift ca. 1980 was attributed to aggravated nutrient loading. Our analysis also shows interactions between these two stressors. Compared to the dynamics driven by nutrient loading alone, WLC reduced the critical P loading and resulted in earlier disappearance of submerged vegetation and emergence of algal blooms by approximately 26 and 10 years, respectively. Overall, our study reveals the significant role of hydrological regulation in driving shallow lake ecosystem dynamics, and it highlights the urgency of using multi-objective management criteria that includes ecological sustainability perspectives when implementing hydrological regulation for aquatic ecosystems around the globe.

Involvement of NADH Oxidase in Competition and Endocarditis Virulence in Streptococcus sanguinis.

Here, we report for the first time that the Streptococcus sanguinis nox gene encoding NADH oxidase is involved in both competition with Streptococcus mutans and virulence for infective endocarditis. An S. sanguinis nox mutant was found to fail to inhibit the growth of Streptococcus mutans under microaerobic conditions. In the presence of oxygen, the recombinant Nox protein of S. sanguinis could reduce oxygen to water and oxidize NADH to NAD(+) The oxidation of NADH to NAD(+) was diminished in the nox mutant. The nox mutant exhibited decreased levels of extracellular H2O2; however, the intracellular level of H2O2 in the mutant was increased. Furthermore, the virulence of the nox mutant was attenuated in a rabbit endocarditis model. The nox mutant also was shown to be more sensitive to blood killing, oxidative and acid stresses, and reduced growth in serum. Thus, NADH oxidase contributes to multiple phenotypes related to competitiveness in the oral cavity and systemic virulence.

Groundwater contamination by microcystin from toxic cyanobacteria blooms in Lake Chaohu, China.

Lake Chaohu is a eutrophic lake that experiences massive cyanobacterial blooms. The high concentrations of microcystin observed in this lake are the result of the bloom's high proportion of toxic cyanobacteria strains. Groundwater is the important source of water for drinking, washing, and irrigation in the watershed of Lake Chaohu. This study examines the space-time distribution of microcystins and related environmental factors in wells near Lake Chaohu. All collected groundwater samples from the Lake Chaohu region had detectable concentrations of microcystins. The highest concentration of microcystins, 1.07 µg L(-1), occurred in a well hundreds of meters from the western coast of the lake in September. The distance from the lake shore to the well was significantly and positively correlated with the microcystin concentration in the groundwater. Moreover, a correlation analysis shows that the microcystin concentration in the groundwater was positively correlated with the total dissolved phosphorus (TDP) and microcystin concentration of the nearby lake water. Therefore, the microcystin in the groundwater likely originates from penetration by nearby lake water. Our results suggest that the groundwater near Lake Chaohu poses a significant health risk for the local residents when used for drinking water.

Effects of elevated CO2 on dynamics of microcystin-producing and non-microcystin-producing strains during Microcystis blooms.

In an attempt to elucidate the effects of different CO2 concentrations (270, 380, and 750 µL/L) on the competition of microcystin-producing (MC-producing) and non-MC-producing Microcystis strains during dense cyanobacteria blooms, an in situ simulation experiment was conducted in the Meiliang Bay of Lake Taihu in the summer of 2012. The abundance of total Microcystis and MC-producing Microcystis genotypes was quantified based on the 16S rDNA and mcyD gene using real-time PCR. The results showed that atmospheric CO2 elevation would significantly decrease the pH value and increase the dissolved inorganic carbon (DIC) concentration. Changes in CO2 concentration did not show significant influence on the abundance of total Microcystis population. However, CO2 concentrations may be an important factor in determining the subpopulation structure of Microcystis. The enhancement of CO2 concentrations could largely increase the competitive ability of non-MC-producing over MC-producing Microcystis, resulting in a higher proportion of non-MC-producing subpopulation in treatments using high CO2 concentrations. Concurrently, MC concentration in water declined when CO2 concentrations were elevated. Therefore, we concluded that the increase of CO2 concentrations might decrease potential health risks of MC for human and animals in the future.

The dynamics of toxic and nontoxic Microcystis during bloom in the large shallow lake, Lake Taihu, China.

Lake Taihu is a large shallow freshwater lake (surface area 2,338 km(2), mean depth 1.9 m) in China, which has experienced toxic cyanobacterial bloom dominated by Microcystis annually during the last few decades. In the present study, the dynamics of toxic and nontoxic Microcystis in three sampling stations (Meiliang Bay (site N2), Gonghu Bay (site N4), and the lake center area (site S4)) were quantified using quantitative real-time PCR (qPCR) during bloom periods from April to September, 2010. Our data showed that the abundance of toxic Microcystis and the toxic proportion gradually increased from April to August in water samples and reached the peak in August. During the study period, toxic Microcystis genotypes comprised between 26.2 and 64.3, between 4.4 and 22.1, and between 10.4 and 20.6 % of the total Microcystis populations in the three sampling sites, respectively. Correlation analysis suggested that there was a strong positive relationship between total Microcystis, toxic Microcystis and the toxic proportion. Chlorophyll a, total phosphorus, and water temperature were positively correlated with the abundances of total Microcystis and toxic Microcystis. Furthermore, the toxic proportion was positively correlated with total phosphorus (P < 0.05) and water temperature (P < 0.01), showing that global warming together with eutrophication could promote more frequent toxic blooms.

Inhibition of Camellia sinensis (L.) O. Kuntze on Microcystis aeruginosa and isolation of the inhibition factors.

Low concentration of tea (Camellia sinensis (L.) O. Kuntze) was shown to inhibit the growth of the toxic cyanobacterium Microcystis aeruginosa. The inhibition efficiency was 40% at 0.1 g dry tea/L and 90% at 0.2 g/L after a 12-day culture. All varieties of tea used in the test could inhibit Microcystis growth, in which the inhibitory effect of green tea was greater than that of black tea. Antialgal allelochemicals were isolated from tea by solvent extraction, gel-chromatography and high performance liquid chromatography. Two algal-inhibition compounds were identified by liquid chromatography/mass spectrometry as epigallocatechin-3-gallate, epicatechin-3-gallate respectively. These are the main polyphenols in tea that have inhibitory effects on the growth of cyanobacteria. The combined effect of these polyphenols makes tea a promising source of algicide to inhibit the growth of algal blooms.

Growth characteristics of algae during early stages of phytoplankton bloom in Lake Taihu, China.

Three treatments, sediment plus lake water (S+W), sterilized sediment plus lake water (SS+W), and sediment plus filtered lake water (S+FW), were recruited to investigate the growth characteristics of algae during pre-bloom and the importance of algal inocula in the water column and sediment. The results showed that in the water column, biomass of all algae increased in all treatments when recruitment was initiated, whereas this tendency differed among treatments with further increment of temperature. The process of algal growth consisted of two stages: Stage I, the onset of recruitment and Stage II, the subsequent growth of algae. Compared with S+W, in Stage I, SS+W significantly increased the biomass of cyanophytes by 178.70%, and decreased the biomass of non-cyanophytes by 43.40%; In Stage II, SS+W notably stimulated the growth of all algae, thus incurring the occurrence of phytoplankton bloom. Further analyses revealed that both metabolic activity and photochemical activity of algae were enhanced in SS+W, which resulted from the releasing of nutrients from sediment. These results suggest that algal growth in Stage II and algal inocula in the water column can be important factors for the formation of phytoplankton bloom. In addition, possible mechanisms promoting algal recruitment and subsequent growth of algae were explored.

Microcystis genotype succession and related environmental factors in Lake Taihu during cyanobacterial blooms.

From spring to autumn, heavy Microcystis blooms always occur in Lake Taihu, although environmental conditions vary markedly. We speculated that Microcystis genotype succession could play an important role in adaptation to environmental changes and long-term maintenance of the high Microcystis biomass. In this study, we investigated Microcystis genotype succession pattern and the related environmental variables in Lake Taihu during cyanobacterial blooms. Denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction -amplified the genus-specific cpcBA and mcyJ gene fragments was used to monitor the variations of Microcystis genotype and potential microcystin (MC)-producing Microcystis genotype compositions during blooms biweekly in three sites (Meiliang Bay, lake center, and Gonghu Bay) and CANOCO 4.5 for Windows were used for the multivariate statistical analysis of their relationships to environmental variables. DGGE patterns indicated that the number of dominant cpcBA genotype per sample increased from spring to autumn. Principal component analysis ordination plots of DGGE profiles showed clear temporal distribution pattern, but not spatial distribution pattern based on both cpcBA and mcyJ genotype compositions. These results indicated there were relatively gradual successions of Microcystis cpcBA and mcyJ genotype compositions in each site, and no distinct spatial difference among the three sites. Redundancy analyses of the gel patterns showed that, in all the three sites, three environmental factors (nitrate, pH, and chemical oxygen demand) were correlated significantly to successions of both cpcBA and mcyJ genotypes except for mcyJ genotype in the lake center. Spearman's correlations indicated that the three environmental variables were also strongly correlated with chl a and MC concentrations. These results suggested that the environmental factors affecting succession of Microcystis community composition might also influence the growth of Microcystis and MC production.

Spatial heterogeneity of cyanobacterial communities and genetic variation of microcystis populations within large, shallow eutrophic lakes (Lake Taihu and Lake Chaohu, China).

Cyanobacteria, specifically Microcystis, usually form massive blooms in eutrophic freshwater lakes. Cyanobacterial samples were collected from eight sites of both Lake Taihu and Lake Chaohu in late summer to determine the diversity and distribution pattern of cyanobacteria and Microcystis in large, shallow, entropic lakes with significant spatial heterogeneity and long-term Microcystis bloom. Molecular methods based on denaturing gradient gel electrophoresis and clone library analysis were used. A similar heterogeneous distribution pattern of cyanobacteria in both lakes was observed. Most parts of these two lakes with high trophic level were dominated by Microcystis. However, in the regions with low trophic levels as well as low concentrations of chlorophyll a, Synechococcus occupied a considerable percentage. Different morphospecies and genotypes dominated the bloom-forming Microcystis populations in these two lakes. Microcystis viridis and Microcystis novacekii were dominant in Lake Chaohu, whereas Microcystis flos-aquae was dominant in Lake Taihu. Only 2 of thel3 Microcystis operational taxonomic units were shared between these two lakes. Analysis of molecular variance based on 16S to 23S internal transcribed spacer sequences indicated the significAnt genetic differentiation of Microcystis between these two lakes (F(ST) = 0.19, p < 0.001). However, only 19.46% of the genetic variability was explained by the population variation between lakes, whereas most (80.54%) of the genetic variability occurred within the lakes. Phylogenetic analysis revealed no phylogeographic structure of Microcystis population in these two lakes, as illustrated by their cosmopolitan nature. Our results revealed that spatial heterogeneity within lakes has more impact on the cyanobacterial diversity than geographical isolation in a local scale.

Quantification of microcystin-producing and non-microcystin producing Microcystis populations during the 2009 and 2010 blooms in Lake Taihu using quantitative real-time PCR.

Lake Taihu, a large, shallow hypertrophic freshwater lake in eastern China, has experienced lake-wide toxic cyanobacterial blooms annually during summer season in the past decades. Spatial changes in the abundance of hepatotoxin microcystin-producing and nonmicrocystin producing Microcystis populations were investigated in the lake in August of 2009 and 2010. To monitor the densities of the total Microcystis population and the potential microcystin-producing subpopulation, we used a quantitative real-time PCR assay targeting the phycocyanin intergenic spacer (PC-IGS) and the microcystin synthetase gene (mcyD), respectively. On the basis of quantification by real-time PCR analysis, the abundance of potential toxic Microcystis genotypes and the ratio of the mcyD subpopulation to the total Microcystis varied significantly, from 4.08 x 10(4) to 5.22 x 10(7) copies/mL, from 5.7% to 65.8%, respectively. Correlation analysis showed a strong positive relationship between chlorophyll-a, toxic Microcystis and total Microcystis; the abundance of toxic Microcystis correlated positively with total phosphorus and ortho-phosphate concentrations, but negatively with TN:TP ratio and nitrate concentrations. Meanwhile the proportion of potential toxic genotypes within Microcystis population showed positive correlation with total phosphorus and ortho-phosphate concentrations. Our data suggest that increased phosphorus loading may be a significant factor promoting the occurrence of toxic Microcystis bloom in Lake Taihu.

THE ACCLIMATIVE CHANGES IN PHOTOCHEMISTRY AFTER COLONY FORMATION OF THE CYANOBACTERIA MICROCYSTIS AERUGINOSA1.

Microcystis aeruginosa (Kütz.) Kütz. commonly occurs as single cells at early recruitment but forms large colonies in summer. Colony formation will induce many acclimative changes. In this study, we demonstrated the photochemical changes before and after colony formation. In the laboratory, light curves showed that colonies were more responsive to high light than single cells. The values of the maximal slope of electron transport rate (ETR)-light curve (α), relative maximal electron transport rate (rETRmax ), and onset of light saturation (Ik ) of colonies were significantly higher than those of single cells (P < 0.05), indicating that colonies have higher photosynthetic capability than single cells, especially in high light, where values of rETRmax and Ik of colonies were 2.32 and 2.41 times those of single cells. Moreover, the dark-light experiments showed that colonial cells can more effectively resist darkness damage. In addition, pigments of colonial cells were higher than those of single cells (P < 0.05). The higher pigment contents probably contribute to higher photosynthetic capability. In the field, the inhibition rate of Fv /Fm in single cells increased significantly faster than that of colonies as light increased (P < 0.05), but nonphotochemical quenching (NPQ) value of colonies was higher (32.4%) than that of single cells at noon, which indicated colonial cells can more effectively resist high-light inhibition than single cells (P < 0.05). Polysaccharides of colonies were significantly higher compared to those in unicellular cells (P < 0.05) based on their contents and ultrastructural characteristics. This finding implies that colonies could not effectively decrease photoinhibition by negative buoyancy regulation. In fact, NPQ may be an important mechanism for avoiding photodamage. All of these phenomena can help explain the ecological success of colonial M. aeruginosa in eutrophic water.

Assessing the combination effects of environmental estrogens in fish.

The method on combined effects of environmental estrogens and mixture environmental risk assessment was discussed. Batch tests were conducted to assess the in vivo potency of mixtures of estrogens using plasma vitellogenin concentrations in male crucian carp (Carassius carassius) as the endpoint. A nonlinear regression was determined on the concentration response relationship for the single chemical of 17ß-estradiol (E(2)), 17α-ethynylestradiol (EE(2)), 4-tert-octylphenol (OP), bisphenol A (BPA), and that of the mixed compounds at equipotent concentrations (E(2)-EE(2), E(2)-EE(2)-OP-BPA), the mixture was tested using a fixed-ratio design. On the basis of statistical selection criteria, the best-fit model is chosen individually for each set of data. Furthermore, the bootstrap methodology is applied for constructing confidence intervals for the estimated effect concentrations. The combined effects of the mixture can be predicted using biomathematical models based on the concentration and potency of the individual mixture components. The finding of non-monotonic dose-response relationship and the combined effects can be accurately predicted in whole range of exposure concentration by the reference models, whereas the outcome of simple effect summation with a great deal of indetermination. Results suggested that there can be a risk of mixture effects. The potential impact of components on mixture would depend predominantly on its concentration, the mixture ratio, and its relative potency. Existing environmental risk assessment procedures are limited in their ability to evaluate the combined effects of chemical mixtures, therefore further improvement is needed.

Vitellogenin induction by a mixture of steroidal estrogens in freshwater fishes and relevant risk assessment.

The study method on combined effects of environmental contaminant mixture and ecological risk assessment was discussed. Batch tests were conducted to assess the in vivo potency of binary mixtures of estrogens using plasma vitellogenin concentrations in male crucian carp as the endpoint. The estrogenic potencies of 17beta-estradiol (E(2)) and 17alpha-ethynylestradiol (EE(2)) were determined following 14 day exposure to the individual chemicals and equipotent binary mixtures. A Nonlinear regression was obtained and 95% confidence limits of effect concentration were achieved using the bootstrap method. Concentration-response curve for fixed ratio binary mixtures of E(2) and EE(2) was compared with those for individual chemicals, using the biomathematical models of concentration addition (CA) and independent action (IA). A complete overlap was found for the CA predictions with the 95% confidence interval of the best-fit regression line of the observed responses, and the IA predictions was shown lower than the observations. The observed mixture effects were considerably higher than those of the hormone alone and far exceeded the 95% confidence interval of the estrogen regression lines. The predicted effects of binary mixtures at different mixture ratios indicated that the potential impact of components on mixture would depend predominantly on its concentration, the mixture ratio and its relative potency. Results suggested that E(2) and EE(2) acted together in an additive manner and the combined effects can be accurately predicted in whole range of exposure concentration by the models of CA and IA, the model of CA might be realistic, but more useful for ecological risk assessment.

Seasonal variation of Microcystis in Lake Taihu and its relationships with environmental factors.

In order to monitor the changes of Microcystis along with temporal and spatial variations, seasonal variation of Microcystis in Lake Taihu was investigated by 16S-23S rRNA internal transcribed spacer denaturing gradient gel electrophoresis (16S-23S rRNA-ITS DGGE) and microscopic evaluation. Samples were collected quarterly at four sites (River Mouth, Meiliang Bay, Cross Area, and Lake Center) from August 2006 to April 2007. Results showed that Microcystis dominated total phytoplankton abundance at the four sites in all seasons except winter. The average annual abundance of Microcystis was relatively high at River Mouth and Meiliang Bay, reaching 81.22 x 10(6) and 61.32 x 10(6) cells/L, respectively. For temporal variations, Shannon-Wiener diversity index (H') according to DGGE profile revealed the richness of Microcystis in summer (H' = 1.375 +/- 0.034) and winter (H' = 1.650 +/- 0.032) was lower than that in spring (H' = 2.078 +/- 0.031) and autumn (H' = 2.365 +/- 0.032) (P <0.05). While for spatial variations, the richness of Microcystis at River Mouth (H' = 2.015 +/- 0.074) was higher than at other sites during four seasons (P < 0.01). Very few differences of Microcystis diversity in the same season were observed among the other three sites (P > 0.05). Canonical correspondence analysis (CCA) was performed to elucidate the relationships between Microcystis operational taxonomic units (OTUs) composition and the environmental factors. Results of CCA revealed that temperature was strongly positively correlated with the first axis (r = 0.963), while TSS was negative correlated with the second axis (r = -0.716). Phylogenetic tree based on the sequencing results of target bands on DGGE gel indicated that samples collected in summer and winter constituted two separated clusters.

Phylogenetic diversity and specificity of bacteria associated with Microcystis aeruginosa and other cyanobacteria.

Interactions between bacteria and cyanobacteria have been suggested to have a potential to influence harmful algal bloom dynamics; however, little information on these interactions has been reported. In this study, the bacterial communities associated with five strains of Microcystis aeruginosa, three species of other Microcystis spp., and four representative species of non-Microcystis cyanobacteria were compared. Bacterial 16S rDNA fragments were amplified and separated by denaturing gradient gel electrophoresis (DGGE) followed by DNA sequence analysis. The similarities among bacterial communities associated with these cyanobacteria were compared to the digitized DGGE profiles using the cluster analyses. The bacterial community structure of all cyanobacterial cultures differed. Cluster analysis showed that the similarity values among M. aeruginosa cultures were higher than those of other cyanobacterial cultures. Sequence analysis of DGGE fragments indicated the presence of bacteria including, Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Bacteroidetes and Actinobacteria in the cyanobacterial cultures. Members of the Sphingomonadales were the prevalent group among the Microcystis-associated bacteria. The results provided further evidence for species-specific associations between cyanobacteria and heterotrophic bacteria, which are useful for understanding interactions between Microcystis and their associated bacteria.

The changes of serum testosterone level and hepatic microsome enzyme activity of crucian carp (Carassius carassius) exposed to a sublethal dosage of pentachlorophenol.

Pentachlorophenol (PCP), which was reported to be a typical persistent organic pollutant and environmental endocrine disruptor, may cause threat to aquatic species. In this study, serum testosterone concentration, activity of liver microsome ethoxyresorufin O-deethylase (EROD) and glutathione S-transferases (GST) of crucian carp (Carassius carassius) exposed to PCP for 7 and 15 d, respectively, were examined. The results showed that testosterone concentration was induced remarkably after 7 d (P<0.05), and the testosterone concentrations in 15 d treatment crucian carp were higher than those in 7 d treatment. It was found that there were significant effects on activities of EROD and GST after crucian carp were exposed to PCP for 7 and 15 d (P<0.05), compared to the controls. EROD and GST activities increased with increase in PCP concentration and also with increase in time on exposure. The results indicated that serum testosterone, EROD and GST were sensitive endpoints to PCP. PCP may have endocrine disrupting activities and may affect the reproductive success of this species. It is possible that the changes of hepatic microsome enzyme activities may result in alterations of serum testosterone levels in crucian carp.

Dynamics and drivers of phytoplankton richness and composition along productivity gradient.

The shape of the productivity-richness relationship (PRR) for phytoplankton has been suggested to be unimodal, that is, the richness peaks at intermediate productivity levels. However, the mechanistic understanding for this pattern is still widely debated. In this study, we observed a unimodal PRR within 71 lakes along the Yangtze River encompassing an altitude range of 0-2700m, and an over 2200km distance from the upper reaches to the lower reaches. At low productivity, the competition for resources and regulatory processes jointly affected phytoplankton richness and composition, and their explanatory power depend on the gradient scale of driving factors. The variation of temperature attributing to altitudinal difference explained the majority of the variations of phytoplankton. If the altitude variation in temperature was eliminated, the explanatory power of temperature decreased from 31.7 to 7.6, and the independent effect of each resource and regulatory variable were limited and not decisive. At high productivity, the negative feedback of increased productivity (light limitation) affected the phytoplankton species richness and composition. The light-sensitive species disappeared, low-light-adapted species was retained and the phytoplankton composition gradually became similar with an increase in productivity. The findings contribute to an increased understanding of the mechanisms resulting in a hump-shaped PRR for phytoplankton.

Large buoyant particles dominated by cyanobacterial colonies harbor distinct bacterial communities from small suspended particles and free-living bacteria in the water column.

Worldwide cyanobacterial blooms greatly impair ecosystems in many eutrophic lakes and impact the microbial environment. In particular, large cyanobacterial colonies that are buoyant on the water surface may provide a distinct habitat for bacteria from other small particles that are suspended stably in the water column. To test this hypothesis, bacterial communities (excluding cyanobacteria) attached to large particles dominated by cyanobacterial colonies (>120 µm, LA), small particles (3-36 µm, SA), and free-living bacteria (0.2-3 µm, FL) were investigated monthly for a year in Lake Taihu, China. Results confirmed that the Shannon diversity index of LA was significantly lower than that of FL, which was lower than that of SA. Cytophagia and Alphaproteobacteria were specially enriched in LA. Although samples in each habitat collected during high- (May to November) and low-bloom seasons (December to April) were separated, all samples in LA were clustered and separated from SA and FL, which were also clustered during the same sampling seasons. In addition, the bacterial communities in LA were correlated with nitrate level, whereas FL and SA were correlated with nitrate level and temperature. Mantel analysis revealed that bacterial composition significantly correlated with the cyanobacterial composition in LA and FL but not in SA. These results indicate that LA provides distinct niches to bacteria, whereas the differentiation of bacterial communities in FL and SA is seasonally dependent.

Bacterial community dynamics and functional variation during the long-term decomposition of cyanobacterial blooms in-vitro.

Cyanobacterial blooms drastically influence carbon and nutrient cycling in eutrophic freshwater lakes. To understand the mineralization process of cyanobacteria-derived particulate organic matter (CyanPOM), the aerobic degradation of cyanobacterial blooms dominated by Microcystis sp. was investigated over a 95-day microcosm experiment. Approximately 91%, 95% and 83% of the initial particulate organic carbon (POC), particulate organic nitrogen (PON), and particulate organic phosphorus (POP) were decomposed, respectively. The POC:PON ratio gradually increased from 5.9 to 13.5, whereas the POC:POP ratio gradually decreased from 230.3 to 120. These results indicated that the coupling of POC, PON, and POP changed during the decomposition of CyanPOM. Moreover, approximately 29%, 51% and 46% of POC, PON, and POP were mineralized to dissolved organic carbon, NO3-, and PO43-, respectively. Rhodospirillales (10.9%), Burkholderiales (16.5%), and Verrucomicrobiales (14.3%) dominated during the rapid phase (days 0-21), whereas Sphingomonadales (12.8%), Rhizobiales (11.8%), and Xanthomonadales (36.5%) dominated during the slow phase (days 21-50) of CyanPOM decomposition. Nitrospira (16.6%-32.9%) dominated and NO3- increased during the refractory phase (days 50-95), thus suggesting the occurrence of nitrification. Redundancy analysis revealed that bacterial communities during rapid decomposition were distinct from those during the slow and refractory periods. POC:POP, NH4+, and NO3- were the major driving factors for the patterns of bacterial communities. Furthermore, increase in nitrogen metabolism, methane metabolism, amino acid related enzymes and pyruvate metabolism characterized the functional variation of bacterial communities during degradation. Therefore, CyanPOM is an important nutrient source, and its decomposition level shapes bacterial communities.