Flavobacteria blooms in four eutrophic lakes: linking population dynamics of freshwater bacterioplankton to resource availability

Heterotrophic bacteria are major contributors to biogeochemical cycles and influence water quality. Still, the lack of representative isolates and the few quantitative surveys leave the ecological role and significance of single bacterial populations to be revealed. Here we analyzed the diversity and dynamics of freshwater Flavobacteria populations in four eutrophic temperate lakes. From each lake, clone libraries were constructed using primers specific for either the class Flavobacteria or Bacteria. Sequencing of 194 Flavobacteria clones from 8 libraries revealed a diverse freshwater Flavobacteria community and distinct differences among lakes. Abundance and seasonal dynamics of Flavobacteria were assessed by quantitative PCR with class-specific primers. In parallel, the dynamics of individual populations within the Flavobacteria community were assessed with terminal restriction fragment length polymorphism analysis using identical primers. The contribution of Flavobacteria to the total bacterioplankton community ranged from 0.4 to almost 100% (average, 24%). Blooms where Flavobacteria represented more than 30% of the bacterioplankton were observed at different times in the four lakes. In general, high proportions of Flavobacteria appeared during episodes of high bacterial production. Phylogenetic analyses combined with Flavobacteria community fingerprints suggested dominance of two Flavobacteria lineages. Both drastic alterations in total Flavobacteria and in community composition of this class significantly correlated with bacterial production, emphasizing that resource availability is an important driver of heterotrophic bacterial succession in eutrophic lakes.     

Regulation of bacterial secondary production in two eutrophic lakes and in experimental enclosures

Bacterial cell number and secondary production of bacterioplanktonwere measured during two periods of 14 and 19 days, respectively,in experimental enclosures of lake water (each containing about5–7 m³ and in two Danish, eutrophic lakes. Additions ofphosphate and planktivorous fish to some of the enclosures inducedrapid changes in the phytoplankton biomass, primary productionand release of extracellular organic carbon (EOC). In some ofthe enclosures containing fish, release of EOC from phytoplanktoncon stituted from 36 to 55% of the bacterial secondary productiondetermined by means of [³H]thymidine incorporation into DNA.In others, such a relationship was not found, suggesting thatphytoplankton species-specific release rates of compositionof EOC might play a role in the microbial response. In enclosureswithout fish, released dissolved organic carbon from zooplanktongrazing activities could Sustain     

Activity profiles of bacterioplankton in a eutrophic river

1. The significance of microbial diversity in processing dissolved organic matter (DOM) is largely unknown. We investigated the range of functional diversity in the bacterioplankton from a eutrophic river by profiling extracellular enzyme activities (EEA) and substrate-induced respiration (SIR) patterns.
2. The EEA profiles consisted of assays for 21 hydrolases, measured using fluorogenic substrates arrayed on 96-well microplates. Commercially available BiOLOG^® GN and ECO plates, which contained 120 different substrates, were used for the SIR profiles.
3. The EEA data were more dynamic than the SIR. Five enzymes, leucine aminopeptidase, alkaline phosphatase, alanine aminopeptidase, arginine aminopeptidase and β-glucosidase, showed consistently high activity; ten others were ubiquitous at lower activity levels; the remainder were detected intermittently. The SIR data showed less temporal variability. With one exception (citrate), the 20 substrates that generated the largest responses were all saccharides or their derivatives.
4. The EEA and SIR data did not generally correlate. Both methods were effective for ordinating bacterioplankton although, unlike the SIR, the EEA ordination followed a clear temporal trajectory.
5. Because the SIR profiles are based on a culture response, whilst the EEA profiles measure activity of the extant community, the latter appear to be more directly linked to the mechanics of DOM processing.     

Relationship between bacterial community composition and bottom-up versus top-down variables in four eutrophic shallow lakes

Bacterial community composition was monitored in four shallow eutrophic lakes during one year using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified prokaryotic rDNA genes. Of the four lakes investigated, two were of the clearwater type and had dense stands of submerged macrophytes while two others were of the turbid type characterized by the occurrence of phytoplankton blooms. One turbid and one clearwater lake had high nutrient levels (total phosphorus, >100 micro g liter(-1)) while the other lakes had relatively low nutrient levels (total phosphorus, <100 micro g liter(-1)). For each lake, seasonal changes in the bacterial community were related to bottom-up (resources) and top-down (grazers) variables by using canonical correspondence analysis (CCA). Using an artificial model dataset to which potential sources of error associated with the use of relative band intensities in DGGE analysis were added, we found that preferential amplification of certain rDNA genes over others does not obscure the relationship between bacterial community composition and explanatory variables. Besides, using this artificial dataset as well as our own data, we found a better correlation between bacterial community composition and explanatory variables by using relative band intensities compared to using presence/absence data. While bacterial community composition was related to phytoplankton biomass in the high-nutrient lakes no such relation was found in the low-nutrient lakes, where the bacterial community is probably dependent on other organic matter sources. We used variation partitioning to evaluate top-down regulation of bacterial community composition after bottom-up regulation has been accounted for. Using this approach, we found no evidence for top-down regulation of bacterial community composition in the turbid lakes, while grazing by ciliates and daphnids (Daphnia and Ceriodaphnia) was significantly related to changes in the bacterial community in the clearwater lakes. Our results suggest that in eutrophic shallow lakes, seasonality of bacterial community structure is dependent on the dominant substrate source as well as on the food web structure.     

Seasonal variability of N:P ratios in eutrophic lakes

The ratios of different forms of nitrogen to phosphorus (particulate, total, total dissolved, and dissolved inorganic N : P) from four eutrophic lakes are presented to assess their variability during two consecutive growing seasons. Particulate and total N : P ratios showed the lowest amplitude of fluctuations, while the total inorganic N : P ratios showed the highest. All N : P forms demonstrated substantial variations and seasonal pattern over the growing period April-October. It was the short-lasting spring minima of the N : P ratios, ranging from less than 1: 1 to 6 : 1, which triggered the onset of nitrogen-fixing cyanophyte blooms. Seasonal mean values were as high as 20: 1 to 30: 1 and misleading in assessing nitrogen limitation. The nitrogen fixation rapidly restored the N : P values to normal levels (TN: TP of 15: 1 and over).     

Genetic diversity of cyanobacteria in four eutrophic lakes

Recent studies indicate genetic diversity of cyanobacteria in eutrophic lakes is not represented well by culture collections or morphology. Yet, few studies have investigated genetic richness and evenness of cyanobacteria using culture-independent methods. We compared the genetic structure of cyanobacteria supported by four neighboring eutrophic lakes during the ice-free season. The partial phycobilincpcB/A genes plus intergenic spacer (PC-IGS) was used as a genetic marker.Sequences were phylogeneticallygrouped by maximum likelihood into genotypes representing sub-genera of the major taxa. Genotypes fell into genera commonly observed by microscopy in these lakes including Microcystis, Aphanizomenon, Chroococcus, Anabaena, and Cylindrospermopsis. Only three genotypes were shared among all four lakes, despite significant water flowage between lakes.A Parsimony P-test indicated lakes were significantly (p=0.01) clustered on the maximum likelihood tree. Pairwise differences using Unifrac distance were moderately or not significant. Analysis of molecular variance (AMOVA) indicated genetic variation among all genotypes (φ=0.06, p<0.001) and 94% of variability occurred within lakes rather than between lakes (6%), explaining the lack of pairwise differences between lakes. Lorenze curves of genotype abundance in each lake showed genetic structure was only moderately uneven (Gini coefficients of 0.37-0.5) indicating lakes did not support dominant genotypes. Overall, results from this study suggest diversity of cyanobacteria is shaped by heterogeneity within lakes (temporally or spatially) and relatively even population structures.     

Distribution of viruses and their impact on bacterioplankton in mesotrophic and eutrophic reservoirs

Spatial distribution of planktonic viral particles (virioplankton) and mortality of heterotrophic bacteria caused by viral lysis were studied in the eutrophic Ivan’kovskoe and mesotrophic Uglichskoe reservoirs (the Upper Volga). During the summer peak of phytoplankton, the number of viral particles was higher in the Ivan’kovskoe Reservoir ((55.1 ± 9.5) × 10⁶ ml⁻¹ on average) than in the Uglichskoe Reservoir ((42.9 ± 5.1) × 10⁶ ml⁻¹ on average). The ratio of viral to bacterial abundances ranged from 2.5 to 7.0. The average number of mature phages in infected heterotrophic bacteria varied from 17 to 109 particles/cell. Most of the infected bacterial cells in the Ivan’kovskoe Reservoir were rod-shaped, and, in the Uglichskoe Reservoir, they were mainly vibrio-shaped. In the Ivan’kovskoe Reservoir, from 8.3 to 22.4% of planktonic bacteria were infected by phages, suggesting phage-induced mortality of bacterioplankton equal to 10.5–34.8% (19.1% on average) of the daily bacterial production. In the Uglichskoe Reservoir, from 9.4 to 33.5% of bacteria were phage-infected, suggesting phage-induced bacterial mortality of 13.7–40.2% (23.5% on average) of the daily bacterial production. The obtained results testify to an important role of autochthonous viruses in the regulation of bacterioplankton abundance and production in the reservoirs.     

Causes and consequences of the success of bream in Dutch eutrophic lakes

In the last decennia eutrophication has caused a shift in the species composition of fish communities in Dutch fresh waters. The changes have led to the disappearance of vegetation in lakes and ponds; zooplankton and chironomids are now the most abundant food organisms for fish. In the turbid, open waters bream and pikeperch are the dominant fish species. Only small bream is vulnerable to predation, but because bream grows much faster than the other cyprinids the time span in which the fish is vulnerable is the shortest. The large bream (>20 cm) can coexist with pikeperch since it is not vulnerable to predation and still utilizes the food organisms efficiently. Eutrophication is accelerated if both bream populations are composed of small-sized specimens preventing large-sized zooplankton to develop, and if they are composed of large-sized individuals which can efficiently stir up the bottom sediments.     

Nutrient limitation of bacterioplankton and phytoplankton in humic lakes in northern Sweden

1. Two small humic lakes in northern Sweden with concentrations of dissolved organic carbon (DOC) between 15 and 20 mg L^–1 were fertilized with inorganic phosphorus (P) and inorganic nitrogen (N), respectively. A third lake was unfertilized and served as a control. In addition to this lake fertilization experiment, data from different regional surveys were used to assess the role of different limiting factors.
2. The P fertilization had no effects on bacterioplankton or phytoplankton, while phytoplankton were significantly stimulated by N fertilization. Inorganic nutrient limitation of bacterioplankton was a function of DOC concentration in water of the investigated region and nutrient-limited bacteria were found only in lakes with DOC concentrations less than around 15 mg L^–1
3. The fertilization experiments demonstrated that the DOC-rich experimental lakes contained a bioavailable pool of P that was not utilized to its full potential under natural conditions. The overall mobilization of energy (bacterioplankton plus phytoplankton) in the experimental lakes was restricted by lack of inorganic N.     

Seasonal modulation of bacterioplankton community at a temperate eutrophic shallow lake

At the present time, there is still a lack of information about environmental parameters modulating variations on bacterial diversity in temperate lakes, particularly from Portugal. Fermentelos Lake (Central Portugal) is a shallow water body that sustains an important wetland area. The strong nutrient inputs from agriculture and industrial runoffs have led to its current eutrophic status. The present work aimed to understand which factors modulate the seasonal bacterioplankton diversity at this lake using 16S rRNA PCR-denaturing gradient gel electrophoresis (DGGE) and multivariate analysis. Environmental data demonstrated eutrophic features throughout all samples with nitrate concentrations reaching 12.0 mg N (NO₃ ⁻) l⁻¹ in March 2006, while the highest conductivity (609 μS cm⁻¹), soluble reactive phosphorus (0.37 mg l⁻¹), total suspended solids (87.2 mg l⁻¹) and chlorophyll a (286.6 μg l⁻¹) levels were recorded in August 2007. Over the past two decades there was a general increase in nitrate, pH and conductivity levels at this lake, suggesting the eutrophication process is still in progress. Multivariate analysis showed that summer versus winter DGGE patterns could be established for bacterial assemblages and were mainly defined by water temperature and chlorophyll a. Actinobacteria were dominant throughout the study period although a general preference for higher temperature, pH, total suspended solids, conductivity, soluble reactive phosphorus (SRP) and chlorophyll a levels was observed. The highest concentrations of nitrogen sources were related to Bacteroidetes and phototrophic eukaryote (cryptophycean) dominance. The expansion of Betaproteobacteria, Alphaproteobacteria and Cyanobacteria phylotypes was generally associated to high temperature, pH, conductivity and SRP values.     

A method to study bacterioplankton community structure in Antarctic lakes

A method to study bacterioplankton community structure in Antarctic freshwater lake samples is described. Small samples (between 300 and 1000 ml) taken in remote field locations were used for crude DNA extraction, followed by PCR amplification of 16S rRNA gene fragments using group-specific primers. The amplification products of the PCR reaction were then separated using denaturing gradient gel electrophoresis to produce a profile of the bacterioplankton community. Whilst the technique is only semi-quantitative, it readily differentiated communities from lakes of different trophic status and from vertical profiles within different lake types. The method offers a sensitive tool for screening and monitoring Antarctic freshwater environments as a precursor and adjunct to more detailed studies. Accepted: 14 November 1999     

Net heterotrophy in Faroe Islands clear-water lakes: causes and consequences for bacterioplankton and phytoplankton

1. Five oligotrophic clear‐water lakes on the Faroe Islands were studied during August 2000. Algal and bacterial production rates, community respiration, and CO₂ saturation were determined. In addition, we examined the plankton community composition (phytoplankton and heterotrophic nanoflagellates) and measured the grazing pressure exerted by common mixotrophic species on bacteria. 2. High respiration to primary production (6.6–33.2) and supersaturation of CO₂ (830–2140 μatm) implied that the lakes were net heterotrophic and that the pelagic heterotrophic plankton were subsidised by allochthonous organic carbon. However, in spite of the apparent high level of net heterotrophy, primary production exceeded bacterial production and the food base for higher trophic levels appeared to be mainly autotrophic. 3. We suggest that the observed net heterotrophy in these lakes was a result of the oligotrophic conditions and hence low primary production in combination with an input of allochthonous C with a relatively high availability. 4. Mixotrophic phytoplankton (Cryptomonas spp., Dinobryon spp. and flagellates cf. Ochromonas spp.) constituted a large percentage of the plankton community (17–83%), possibly as a result of their capacity to exploit bacteria as a means of acquiring nutrients in these nutrient poor systems.     

Effect of humic material on the bacterioplankton community composition in boreal lakes and mesocosms

The bacterioplankton community composition in two Finnish forest lakes with different content of humic substances was studied by denaturing gradient gel electrophoresis (DGGE) and sequencing of the major bands. The same dominant bacterial phylotypes were detected in the bacterioplankton communities of clear-water Lake Ahvenlammi and humic Lake Sammalisto. For 4 years, in every water layer, Actinobacteria was the dominant and Verrucomicrobia the second most common phylum. In the hypolimnion, other dominant phyla were also found. We set up a mesocosm experiment to assess the effect of a sudden load of allochthonous humus extract to the bacterioplankton community composition. Changes in the bacterial communities were followed in four control and four humus extract-added mesocosms for 50 days. In the humic mesocosms the phylotypes of allochthonous Proteobacteria arriving with the humus extract were initially prevalent but disappeared during the first weeks. After this the Actinobacteria-dominated communities resembled the bacterioplankton communities of the control mesocosms and Lake Ahvenlammi. Towards the end of the experiment the community patterns in all the mesocosms started to change slightly because of erratic occurrence of new proteobacterial phylotypes. Thus the effects of a sudden load of allochthonous humic material and bacteria to the bacterioplankton community composition were transient.     

Influence of hydrological seasonality on bacterioplankton in two neotropical floodplain lakes

Seasonal fluctuation in river stage strongly affects the ecological functioning of tropical floodplain lakes. This study was conducted to assess the influence of hydrological seasonality on bacterial production and abundance in two floodplain lakes of the Autana River, a blackwater river in the Middle Orinoco basin, Venezuela. Water samples for nutrient chemistry, chlorophyll a, and microbiological determinations were collected in two floodplain lakes and in the mainstem of the river during 1997–98. DOC and chlorophyll a concentrations were similar between mainstem and lake sites during high water when river and lakes were well connected but became different during the period of low water when the interaction was minimal. Higher values of bacterial production were observed in the floodplain lakes (0.62–1.03 g C l^–1 h^–1) compared to the mainstem sites (0.17–0.19 g C l^–1 h^–1) during the period of low water, while during the period of high water river and lake sites showed similar levels (0.04 g C l^–1 h^–1). Bacterial numbers followed bacterial production in the floodplain lakes, reaching higher numbers during the period of low water (1.41–2.40 × 10⁶ cells ml^–1). Availability of substrate and inorganic nutrients, pH, and inputs and losses of bacterial cells could be determining the observed seasonal patterns in bacterial production and abundance. The Autana lakes exhibited a strong seasonal pattern in the chemical and biological conditions, showing higher productivity during the lentic phase that lasted between 5 and 6 months.     

Uptake of amino acids by uncultivated bacterioplankton of a eutrophic water reservoir

Water bacterioplankton species (clones) feeding on lysine and glycine were revealed in experimental microecosystems using nonsterile cultivation. The structure of the community was analyzed by separation of amplified fragments of the bacterial 16S rRNA gene of by denaturing gradient gel electrophoresis. The dominant species (clones) of the bacterioplankton were identified by molecular cloning and determining the nucleotide sequence of part of the 16S gene of ribosomal RNA. It was found that glycine, one of the prevalent amino acids of a eutrophic reservoir, and lysine, whose content in natural water is lower, are nutrient substrates for various subdominant bacterioplankton species (clones). Thus, free-living bacterial species are highly specialized for the uptake of individual amino acids.     

Submerged vegetation development in two shallow, eutrophic lakes

Submerged macrophyte vegetation in two shallow lakes in the Netherlands, Lake Veluwemeer and Lake Wolderwijd, has been affected by eutrophication in the late 1960's and 1970's. Recent changes in the vegetation occurred in the period following lake restoration measures. Between 1987 and 1993, the dominance of Potamogeton pectinatus decreased, while Charophyte meadows expanded over the same time interval. The pattern of change of the dominant macrophyte species might result from changes in the underwater light climate. Seasonally persistent clear water patches associated with the Chara meadows have been observed in the last few years. The interaction between submerged macrophyte vegetation succession and water transparency in the lakes is discussed.     

水动力扰动方式对富营养化湖泊藻毒素的影响

太湖是一个典型的大型浅水湖泊,在季风影响下,常年受风浪扰动影响。为了解风浪扰动对太湖水体中微囊藻释放藻毒素(MC)的影响,于2018年7月采集含有微囊藻水华的太湖原水,在中国科学院太湖湖泊生态系统研究站设置了对照、间歇扰动和持续扰动3种处理方式来进行模拟实验,研究水体中藻毒素的变化,实验共持续19 d。结果表明:3种处理方式下,MC浓度的变化趋势一致,1～13 d呈升高趋势,其后均有不同幅度的下降;对照组MC浓度最低,平均值为1.69μg·L-1,显著低于间歇扰动组和持续扰动组(P<0.05);间歇扰动组和持续扰动组MC均值分别为1.81和1.86μg·L-1,两者差异不显著; 3种处理方式下,单位藻细胞MC含量1～7 d持续减小,且各组间差异不显著(P>0.05); 10～19 d各组均有一个先增加后减小的过程,且组间差异均显著(P<0.05),对照组、间歇扰动组和持续扰动组单位藻细胞MC含量均值分别为0.55、1.20和1.98μg·108cells-1;本实验条件下MC与氮磷等因子显著负相关,与水温、SS、Chla等之间显著正相关。本研究表明,扰动可促进水体中藻...     

Mechanisms regulating abundance of submerged vegetation in shallow eutrophic lakes

Shallow eutrophic lakes tend to be either in a turbid state dominated by phytoplankton or in a clear-water state dominated by submerged macrovegetation. Recent studies suggest that the low water turbidity in the clear-water state is maintained through direct and indirect effects of the submerged vegetation. This study examined what mechanisms may cause a recession of the submerged vegetation in the clear-water state, and thereby a switch to the turbid state. The spatial distribution of submerged vegetation biomass was investigated in two shallow eutrophic lakes in the clear-water state in southern Sweden. Biomass of submerged vegetation was positively correlated with water depth and wave exposure, which also were mutually correlated, suggesting that mechanisms hampering submerged vegetation were strongest at shallow and/or sheltered locations. The growth of Myriophyllum spicatum, planted in the same substrate and at the same water depth, was compared between sheltered and wave exposed sites in two lakes. After 6 weeks the plants were significantly smaller at the sheltered sites, where periphyton production was about 5 times higher than at the exposed sites. Exclosure experiments were conducted to evaluate the effects of waterfowl grazing on macrophyte biomass. Potamogeton pectinatus growth was decreased by grazing, whereas M. spicatum was not affected. The effects were greater at a sheltered than at a wave-exposed site, and also negatively related to distance from the reed belt. These results suggest that competition from epiphytes and waterfowl grazing hamper the development of submerged vegetation at sheltered and/or shallow locations. An increased strength of these mechanisms may cause a recession of submerged vegetation in shallow eutrophic lakes in the clear-water state and thereby a switch to the turbid state. Received: 24 June 1996 / Accepted: 8 September 1996     

Respiration of plankton in two small,polyhumic lakes

The respiration of plankton of two polyhumic lakes was measured as production of carbon dioxide in dark bottles. The method proved to be enough sensitive for use in oligotrophic lakes with low alkalinity. The respiration of plankton followed broadly changes in temperature. However, the primary production of phytoplankton was probably the main factor governing the seasonal pattern of respiration. During summer the respiration of plankton was more than three times higher than the primary production of phytoplankton. This suggests that allochthonous humic substances are an important source of carbon and energy for organisms of polyhumic lakes.