Seasonal Periodicity in Lacustrine Phytoplankton and the Theory of Ecological Succession

An attempt has been made to examine the extent to which seasonal periodicity of lacustrine phytoplankton corresponds to the general concepts of the theory of ecological succession. The study was carried out on an eutrophic lake (Aydat, France). As REYNOLDS (1980, 1984) suggests, the seasonal periodicity of phytoplankton is an autogenic succession which may be perturbed by allogenic physical processes that change the initial direction of the succession. Thus, in this lake, only spring succession of phytoplankton can be considered as a true succession, consistent with the theory of ecological succession in that it was a directional and predictable phenomenon leading to an increasing complexity of community.     

Seasonal succession and strategies of phytoplankton development in two lakes of different trophic states

This study describes the metabolism and structure of phytoplanktoncommunities during seasonal periodicity and discusses strategiesof development adopted by species during succession. The studywas conducted in two trophically different lakes. Each lakedemonstrates a different degree of the ecological succession,which prescribes an increasingly complex taxocenose. In oligomesotrophicLake Pavin the autogenic succession lasts from spring overturnuntil mid-summer. In eutrophic Lake Aydat the autogenic successionis strictly limited to spring. The seasonal changes of the communityproductivity, turnover rate (P/B) and adenylate energy chargeconfirm the hypothesis of a change of the ‘metabolic orientation’of phytoplankton cells during seasonal succession. The autogenicsuccession represents the progression from a growth-orientedstrategy to an equilibrium-oriented one.     

Exploring and quantifying fungal diversity in freshwater lake ecosystems using rDNA cloning/sequencing and SSU tag pyrosequencing

Water samples were collected along transects from the shore to the centre of two French lakes: the deep, volcanic, oligomesotrophic and low allochthonic-impacted Lake Pavin, and the productive and higher allochthonic-impacted Lake Aydat. The biodiversity was analysed using two approaches: the classical approach consisting of cloning/sequencing of the 18S, ITS1, 5.8S, ITS2 and partial 28S region using primers designed for fungus sequences, and the pyrosequencing of 18S rRNA hypervariable V2, V3 and V5 regions using two primer sets (one universal for eukaryotes and one for fungi). The classical approach yielded 146 (Lake Pavin) and 143 (Lake Aydat) sequences, corresponding to 46 and 63 operational taxonomic units (OTUs) respectively. Fungi represented half of the OTUs identified in Lake Pavin and 30% in Lake Aydat, and were dominated by sequences from Chytridiomycota found throughout Lake Pavin but mostly in the central pelagic zone of Lake Aydat. The pyrosequencing approach yielded 42,064 (Pavin) and 61,371 (Aydat) reads, of which 12-15% and 9-19% reads were assigned to fungi in Lakes Pavin and Aydat respectively. Chytridiomycota members were also dominant among these reads, with OTUs displaying up to > 33-fold overrepresentation in the centre compared with the riparian areas of Lake Aydat. Besides fungi, both approaches revealed other major eukaryote groups, with the highest diversity in the central areas of lakes. One of the major findings of our study was that the two lakes displayed contrasting spatial distributions, homogenous for Lake Pavin and heterogeneous for Lake Aydat, which may be related to their peculiarities. This study represents the first unveiling of microbial eukaryote and fungus diversity assessed with two complementary molecular methods, and is considered a major milestone towards understanding the dynamics and ecology of fungi in freshwater lake ecosystems, which are directly link to the abundance and distribution of taxa.     

Viral activity in two contrasting lake ecosystems

For aquatic systems, especially freshwaters, there is little data on the long-term (i.e., >6-month period) and depth-related variability of viruses. In this study, we examined virus-induced mortality of heterotrophic bacteria over a 10-month period and throughout the water column in two lakes of the French Massif Central, the oligomesotrophic Lake Pavin and the eutrophic Lake Aydat. Concurrently, we estimated nonviral mortality through heterotrophic nanoflagellate and ciliate bacterivory. Overall, viral infection parameters were much less variable than bacterial production. We found that the frequency of visibly infected cells (FVIC), estimated using transmission electron microscopy, peaked in both lakes at the end of spring (May to June) and in early autumn (September to October). FVIC values were significantly higher in Lake Pavin (mean [M] = 1.6%) than in Lake Aydat (M = 1.1%), whereas the opposite trend was observed for burst sizes, which averaged 25.7 and 30.2 virus particles bacterium(-1), respectively. We detected no significant depth-related differences in FVIC or burst size. We found that in both lakes the removal of bacterial production by flagellate grazing (M(Pavin) = 37.7%, M(Aydat) = 18.5%) was nearly always more than the production removed by viral lysis (M(Pavin) = 16.2%, M(Aydat) = 19%) or ciliate grazing (M(Pavin) = 2.7%, M(Aydat) = 8.8%). However, at specific times and locations, viral lysis prevailed over protistan grazing, for example, in the anoxic hypolimnion of Lake Aydat. In addition, viral mortality represented a relatively constant mortality source in a bacterial community showing large variations in growth rate and subject to large variations in loss rates from grazers. Finally, although viruses did not represent the main agent of bacterial mortality, our data seem to show that their relative importance was higher in the less productive system.     

Virioplankton and microbial communities in aquatic systems: a seasonal study in two lakes of differing trophy

1. The seasonal and vertical distribution of the abundance of virus‐like particles (VLPs), together with the abundances of other microbial organisms (bacteria, unpigmented and pigmented nanoflagellates and ciliates), were determined in an oligomesotrophic lake (Pavin, France) and in a eutrophic lake (Aydat, France) between March and December 2000. 2. The abundance of the viral plankton and those of other microbial taxa were significantly higher in the more productive system. The same was for the virus‐to‐bacteria quotient (VBQ), which averaged seven in Lake Pavin and nine in Lake Aydat. 3. The abundance of viruses increased during the period of thermal stratification in both lakes, with the highest values being recorded at the end of summer/early autumn in the epi‐ and the metalimnion. The seasonal pattern of abundance of viruses in both lakes in the surface layer was similar, indicating that the dynamics of viruses may be controlled by environmental factors such as light conditions. 4. There was no correlation between the abundance of viruses and protists. We found correlations between viruses and heterotrophic bacteria in the whole water column in Lake Pavin, but only in the dark bottom waters in Lake Aydat. 5. Overall, the empirical findings in this study lead us to speculate that the weaker correlation between bacteria and viruses in Lake Aydat than in Lake Pavin, as well as the higher VBQ in the former, is a consequence of the increasing relative abundance of non‐bacteriophage VLPs along the trophic gradient of aquatic systems.     

Grazing and assimilation rates of natural populations of planktonic rotifersKeratella cochlearis,Keratella quadrata andKellicottia longispina in a eutrophic lake (Aydat,France)

The filtering rates of¹⁴C labelledChlamydomonas sp. by 3 dominant species of rotifers were studied in eutrophic Lake Aydat. They varied from 4 to 53 µl ind^–1 h^–1 forKeratella cochlearis, from 2 to 56 µl.^–1h^–1 forKeratella quadrata and from 3 to 52 µl ind^–1 h^–1 forKellicottia longispina. Their maximum assimilation efficiency was 32%. At the measured grazing rates, these populations could clear the water in less than two days during July. In Lake Aydat, the rotifers community could play an important role in the regulation of seasonal succession of phytoplankton and bacteria.     

8 Grazing by heterotrophic nanoflagellates on virus- and bacteria-sized particles in two lakes of different trophy

Although viruses are now widely recognized to infect aquatic bacteria as well as prokaryotic and eukaryotic phytoplankton, our understanding of how they fit into aquatic foodwebs remains still deficient. The present study examined the potential nutritional food source that viral particles represent for natural assemblages of heterotrophic nanoflagellates (HNF). We determined the seasonal and depth-related variability in the grazing activity of HNF on virus- and bacteria-sized particles in the oligomesotrophic Lake Pavin and the eutrophic Lake Aydat. Ingestion rates were determined using 50 and 500 nm diameter fluorescent microspheres. Estimated ingestion rates ranged from 0.01 to 1.7 viruses cell⁻¹·h⁻¹ (Aydat) and from 0.01 to 2 viruses cell⁻¹·h⁻¹ (Pavin). Derived clearance rates for viruses represented 3–5% (Aydat) and 10–12% (Pavin) of those for bacteria. In general, HNF grazing on both viruses and bacteria showed a similar pattern characterized by an apparent decrease in summer followed by autumnal peaks, with lowest values always being recorded in the hypolimnion. Viral production, estimated from the frequency of visibly infected cells (using transmission electronic microscopy), was consumed by HNF at rates averaging 2% (Aydat) and 10% (Pavin). This study suggests that, viruses, which are assumed to correspond to 1×10⁻⁷ ng C/particle, are likely to be of low nutritive value for HNF in both lakes. However, these results imply that HNF grazing may contribute significantly to the removal of viruses in freshwaters, especially in low productive systems.     

Genetic diversity of small eukaryotes in lakes differing by their trophic status

Small eukaryotes, cells with a diameter of less than 5 mum, are fundamental components of lacustrine planktonic systems. In this study, small-eukaryote diversity was determined by sequencing cloned 18S rRNA genes in three libraries from lakes of differing trophic status in the Massif Central, France: the oligotrophic Lake Godivelle, the oligomesotrophic Lake Pavin, and the eutrophic Lake Aydat. This analysis shows that the least diversified library was in the eutrophic lake (12 operational taxonomic units [OTUs]) and the most diversified was in the oligomesotrophic lake (26 OTUs). Certain groups were present in at least two ecosystems, while the others were specific to one lake on the sampling date. Cryptophyta, Chrysophyceae, and the strictly heterotrophic eukaryotes, Ciliophora and fungi, were identified in the three libraries. Among the small eukaryotes found only in two lakes, Choanoflagellida and environmental sequences (LKM11) were not detected in the eutrophic system whereas Cercozoa were confined to the oligomesotrophic and eutrophic lakes. Three OTUs, linked to the Perkinsozoa, were detected only in the Aydat library, where they represented 60% of the clones of the library. Chlorophyta and Haptophyta lineages were represented by a single clone and were present only in Godivelle and Pavin, respectively. Of the 127 clones studied, classical pigmented organisms (autotrophs and mixotrophs) represented only a low proportion regardless of the library's origin. This study shows that the small-eukaryote community composition may differ as a function of trophic status; certain lineages could be detected only in a single ecosystem.     

Seasonal variations of phytoplankton photosynthate partitioning in two lakes of different trophic level

The aim of this study was to compare vertical and seasonal variationsof inorganic carbon allocation into macromolecules by the phytoplanktonpopulation in a eutrophic lake (Lake Aydat) and an oligo-mesotrophiclake (Lake Pavin). Biochemical fractionation was conducted byconsecu tive differential extractions in order to separate proteins,polysaccharides, lipids, and low molecular weight compounds(LMW). The ratio of light absorption at480 and 665 nm by acetoneextracts of phytoplankton pigments was used as an indicatorof the nutritional statusof natural phytoplankton populations.Our results show that in Lake Aydat, the main photosyntheticend productswere poly saccharides, whereas in Lake Pavin, radioactivitywas predominantly incorporated into the protein fraction. Moreover,the seasonal cycles of mixing and stratification in these twolakes affected the pattern of ¹⁴C incorporation into LMW andmacromolecules. An increase in the relative synthesis of proteinsoccurred during stratification periods. It was linked to anincrease in temperature and nutrient limitation further complicatedby the shift in species composition of the populations. Differences recorded both between the two lakes of different trophicstatus and between seasons confirm that the proportion of carbonincorporated into proteins might be a useful indicator of thephysio logical status of phvtoplankton communities.     

Phytoplankton chytridiomycosis: community structure and infectivity of fungal parasites in aquatic ecosystems

Fungal parasitism is recurrent in plankton communities, especially in the form of parasitic chytrids. However, few attempts have been made to study the community structure and activity of parasites at the natural community level. To analyse the dynamics of zoosporic fungal parasites (i.e. chytrids) of phytoplankton, samples were collected from February to December 2007 in two freshwater lakes. Infective chytrids were omnipresent in lakes, with higher diversity of parasites and infected phytoplankton than in previous studies. The abundance and biomass of parasites were significantly higher in the productive Lake Aydat than in the oligomesotrophic Lake Pavin, while the infection prevalence in both lakes were similar and averaged about 20%. The host species composition and their size appeared as critical for chytrid infectivity, the larger hosts being more vulnerable, including pennate diatoms and desmids in both lakes. The highest prevalence (98%) was noted for the autumn bloom of the cyanobacterium Anabaena flosaquae facing the parasite Rhizosiphon crassum in Lake Aydat. Because parasites killed their hosts, this implies that cyanobacterial blooms, and other large size inedible phytoplankton blooms as well, may not totally represent trophic bottlenecks because their zoosporic parasites can release dissolved substrates for microbial processes through host destruction, and provide energetic particles as zoospores for grazers. Overall, we conclude that the parasitism by zoosporic fungi represents an important ecological driving force in the food web dynamics of aquatic ecosystems, and infer general empirical models on chytrid seasonality and trophodynamics in lakes.     

Grazing and assimilation rates of natural populations of planktonic cladocerans in a eutrophic lake

The filtering rates of ¹⁴ carbon-labelled Chlamydomonas sp. by 4 dominant species of Cladocera were studied in eutrophic Lake Aydat. The mean individual filtering rates were 364, 399, 403 and 408 µl ind^–1 h^–1 for Bosmina longirostris, Daphnia longispina, Chydorus sphaericus and Ceriodaphnia quadrangula, respectively. Their maximum assimilation efficiencies attained ± 50%. These populations collectively could consume the available food in less than 4 days during August (maximum daily grazing rate = 29%).In eutrophic conditions lake Aydat D. longispina appears to be more efficient in energy utilization than other cladocerans. Some individuals of Daphnia and Chydorus continued to filter in the zone of low oxygen where an important bacterial biomass had developed. Our results support the importance of the cladocerans in the regulation of phytoplankton production.     

Rapid quantification of planktonic ciliates: comparison of improved live counting with other methods

THE FOLLOWING EFFICIENT AND QUANTITATIVELY VALID METHOD TO FILTER CONCENTRATE AND COUNT LIVE PLANKTONIC CILIATES WAS DEVELOPED AND COMPARED WITH OTHER TREATMENTS: unconcentrated (raw) samples and centrifuged samples were counted live, and the effects of five different fixatives (HgCl(2), Lugol's iodine, formaldehyde, glutaraldehyde, and Champy-DaFano) on the counts were monitored. Samples originated from a eutrophic mountain lake (Lake Aydat, near Clermont-Ferrand, France). Overall, live filtered counts were similar to counts of raw samples, but they were significantly higher (2 to 2.3 fold, P < 0.05) by analysis of variance than counts from centrifuged samples. Nevertheless, some taxa, i.e., Halteria and Loxodes spp., were sensitive to filtration. The live filtered counts were also comparable to counts of raw HgCl(2)-fixed and settled samples. HgCl(2) and Lugol fixation consistently gave the highest total counts, while significantly lower counts were always obtained with Champy-DaFano-fixed samples. Losses due to fixation were insignificant for raw samples but were substantial and statistically significant in concentrated samples (15% after filtration and 71% after centrifugation, compared with counts from the corresponding live samples). Live counting of passively filter-concentrated ciliates has many advantages over other methods. It is two to four times quicker and more efficient. Ciliates are recognized with certainty, more species are identified, and enumeration of dead organisms (e.g., tintinnid loricas) is avoided. It should be recommended as a quantitatively valid alternative to classical methods for assessing planktonic ciliate populations.     

Viral Activity in Two Contrasting Lake Ecosystems

For aquatic systems, especially freshwaters, there is little data on the long-term (i.e., >6-month period) and depth-related variability of viruses. In this study, we examined virus-induced mortality of heterotrophic bacteria over a 10-month period and throughout the water column in two lakes of the French Massif Central, the oligomesotrophic Lake Pavin and the eutrophic Lake Aydat. Concurrently, we estimated nonviral mortality through heterotrophic nanoflagellate and ciliate bacterivory. Overall, viral infection parameters were much less variable than bacterial production. We found that the frequency of visibly infected cells (FVIC), estimated using transmission electron microscopy, peaked in both lakes at the end of spring (May to June) and in early autumn (September to October). FVIC values were significantly higher in Lake Pavin (mean [M] = 1.6%) than in Lake Aydat (M = 1.1%), whereas the opposite trend was observed for burst sizes, which averaged 25.7 and 30.2 virus particles bacterium⁻¹, respectively. We detected no significant depth-related differences in FVIC or burst size. We found that in both lakes the removal of bacterial production by flagellate grazing (M_Pavin = 37.7%, M_Aydat = 18.5%) was nearly always more than the production removed by viral lysis (M_Pavin = 16.2%, M_Aydat = 19%) or ciliate grazing (M_Pavin = 2.7%, M_Aydat = 8.8%). However, at specific times and locations, viral lysis prevailed over protistan grazing, for example, in the anoxic hypolimnion of Lake Aydat. In addition, viral mortality represented a relatively constant mortality source in a bacterial community showing large variations in growth rate and subject to large variations in loss rates from grazers. Finally, although viruses did not represent the main agent of bacterial mortality, our data seem to show that their relative importance was higher in the less productive system.     

Effects of nitrogen nutritional deficiency on the distribution of particular lipids in a eutrophic lake milieu]

The seasonal distribution of particulate lipids coupled with N-nutrient availability was studied in eutrophic Lake Aydat (Massif Central, France). The concentrations of lipids ranged between 196.9 and 2971.5 micrograms.l-1 (mean +/- s.d. = 1090.1 +/- 705.5 micrograms.l-1. Lipids were abundant in summer and fall when nitrates were insufficient reflecting thus an orientation of cell metabolism towards an accumulation of storage products. In such conditions, the heterocystous Cyanobacteria were found to develop due to their competitive advantage of exploiting atmospheric nitrogen. Their lipid metabolism did not seem to be affected at least partially by NO3- exhaustion.     

High lytic infection rates but low abundances of prokaryote viruses in a humic lake (Vassivière, Massif Central, France)

We explored the abundance and infection rates of viruses on a time series scale in the euphotic zone of the humic mesotrophic Lake Vassivière (Massif Central, France) and compared them to nonhumic lakes of contrasting trophy (i.e., the oligomesotrophic Lake Pavin and the eutrophic Lake Aydat) located in the same geographical region and sampled during the same period. In Lake Vassivière, the abundances of virus-like particles (range, 1.7 × 10(10) to 2.6 × 10(10) liter(-1)) were significantly (P < 0.001) lower than in Lakes Pavin and Aydat. The percentage of virus-infected prokaryotic cells (mean, 18.0%) was significantly higher (P < 0.001) in Vassivière than in Pavin (mean, 11.5%) and Aydat (mean, 9.7%). In Vassivière, the abundance of prokaryotes was a good predictor (r = 0.78, P < 0.001) of the number of virus-like particles, while the potential grazing rate from heterotrophic nanoflagellates was positively correlated to the viral infection rate (r = 0.75, P < 0.001; n = 20), indicating the prevalence of cycling interactions among viruses, prokaryotes, and grazers, which is in agreement with past experiments. The absence of correlation between chlorophyll a concentrations (Chl) and viral parameters suggested that the resources for the lytic activity of viruses in Vassivière were mainly under allochthonous control, through host activity. Indeed, compilation of data obtained from several nonhumic lakes in the French Massif Central revealed that Chl was positively correlated to the abundance of virus-like particles at concentrations above 0.5 μg Chl liter(-1) and negatively at concentrations below 0.5 μg Chl liter(-1), suggesting that phytoplankton-derived resources could force prokaryotic growth to attain a certain threshold level when the host availability is sufficient to boost the proliferation of viruses. Therefore, based on the high level of lytic infection rates in Lake Vassivière, we conclude that viruses are key agents for prokaryotic mortality and could influence the food web dynamics in humic lakes, which may ultimately depend on the internal cycling of resources and, perhaps, mainly on the allochthonous inputs and the associated humic substances.     

Science, Art, or Application—the "Karma" of Restoration Ecology

The present state of restoration ecology is far away from Bradshaw’s “acid test for ecology.” The conclusions drawn from the series of papers in this issue and from the Jena workshop suggest some directions in which the field may progress. More attention must be paid to the degraded state, which should be evaluated by its specific features and carefully analyzed before any restoration plan is laid down. Restoration goals have to be realistic, which includes the appreciation of globally changing conditions, resulting in a paradigm‐shift toward “forward‐restoration.” Basically, the transition from the degraded state conditions to the target state is a kind of succession that is manipulated by the application of goal‐orientated and system‐specific disturbances. Whenever possible, restorationists should step back and make use of naturally occurring succession, which requires a sophisticated restoration strategy, involving flexible management responses, multiple alternative target states, robust measurements for the restoration progress, and careful long‐term monitoring. The unique feature of restoration ecology is the involvement of socioeconomic decisions, and conceptual frameworks for ecological restoration have to implement the specific links to natural succession. To bridge the gap between ecological theory and on the ground restoration, it is essential that restoration practice is translated into the vocabulary and thinking of basic ecology. If all these aspects are integrated, ecological restoration as an application—and restoration ecology as an applied science—may develop into an acid test for our understanding of interactions between people and their environment, rather than for pure ecology.     

Molecular and morphological diversity of fungi and the associated functions in three European nearby lakes

This study presents an original rDNA PCR and microscopic survey of pelagic freshwater fungal communities, and was designed to unveil the diversity of true Fungi (i.e. the kingdom Eumycota) in three contrasting lake ecosystems (Lakes Pavin, Aydat and Vassivière) located in the French Massif Central. Three clone libraries were constructed from samples collected in the euphotic layers of the lakes during spring 2007. Phylogenetic analysis of the combined data from the three lakes clustered our sequences into thee divisions: Chytridiomycota (50% of total sequences), Ascomycota (40%) and Basidiomycota (10% in Pavin and Aydat only). Several sequences were assigned to a novel Chytridiomycota clade first recovered in Lake Pavin in 2005. Most of the sequences retrieved in the investigated lakes were affiliated with known fungal species, most of which were apparently well adapted to thrive in the pelagic realm. Their main functions (i.e. parasitism and saprophytism), putatively inferred from the closest relatives of the retrieved molecular sequences, were confirmed by microscopic approaches and by enrichment experiments with pollen grains. The occurrence of three fungal forms (zoosporic, yeast and mycelial) was associated with different trophic modes, establishing fungi as strong potential competitors for various niches in pelagic ecosystems, primarily in relation to the processing of particulate organic matter and the production of propagule food sources for grazers. For the first time, this study provides insight into the diversity and the associated functions of all members of the Kingdom Eumycota investigated in the whole plankton fraction of aquatic ecosystems.     

Characterization of akinetes from cyanobacterial strains and lake sediment: A study of their resistance and toxic potential

Nostocalean cyanobacteria are known to proliferate abundantly in eutrophic aquatic ecosystems, and to produce several cyanotoxins, including anatoxin-a. In this study, we investigated both the resistance and toxic potential of the akinetes (resistant cells), using cyanobacterial cultures and akinetes extracted from the sediment of Lake Aydat (France) sampled in the winter and spring. Intact and lysed akinetes were differentiated using a double control based on the autofluorescence of akinetes and SYTOX-green staining. The percentage of resistant akinetes found in several different abiotic stress conditions was highly variable, depending on the species and also on the sampling season. Thus, the resistance of akinetes and their ability to germinate seems to follow a species-specific process, and akinetes can undergo physiologic changes during the sedimentary phase of the Nostocale life cycle. This study also revealed the first evidence of anatoxin-a genes in akinetes, with anaC and anaF genes detected in akinetes from all cyanobacterial producer cultures. The low number of anaC genes, almost exclusively detected using nested PCR, in the sediment at Lake Aydat suggests a limited but existent past population of toxic Nostocales in this lake. Given the key role of akinetes in the annual cycle and subsequent summer proliferation, it can be interesting to integrate the surveillance of akinetes in the management of lakes exposed to recurrent cyanobacterial blooms.     

湖湾效应对长江中游湖泊水生植物多样性的影响

在３ａ野外调查和定位研究的基础上,本文对长江中游地区水生植物物种多样性进行了初步研究。主要工作在１０个有代表性的湖泊进行,结果表明：湖泊水生植物多样性指数沿湖湾至中央敞水区由２．１６５５降至０．３６３３;植物种类和个体数自湖湾至湖心区急剧减少,植物群落生物量自湖湾至湖中央区也发生明显变化;水深梯度与植物种类分布密切相关,在保安湖０．５ｍ水深中有１９种植物,在２ｍ水深中仅有４种植物,绘出３个湖的种－