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.     

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.     

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.     

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.     

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.     

Viral lysis,flagellate grazing potential,and bacterial production in Lake Pavin

Abundances of different compartments of the microbial loop (i.e., viruses, heterotrophic bacteria, nonpigmented nanoflagellates, and pigmented nanoflagellates), bacterial heterotrophic production (BHP), viral lysis, and potential flagellate grazing impacts on the bacterial assemblages were estimated during a short-term study (24 h) conducted in June 1998 in the epilimnion (5 m) and metalimnion (10 m) of a moderate-altitude oligomesotrophic lake (Lake Pavin, France). Viral and bacterial abundances were higher in the metalimnion than in the epilimnion, whereas pigmented and nonpigmented nanoflagellates were more numerous in the epilimnion. The control of the BHP due to viral lysis (determined by examination of viral-containing bacteria using a transmission electron microscope) was significantly higher in the meta- (range = 6.0-33.7%, mean = 15.6%) than in the epilimnion (3.5-10.3%, 6.4%). The same was for the losses of BHP from the potential predation by nanoflagellates which ranged from 0.5 to 115.4% (mean = 38.7%) in the epilimnion, and from 0.7 to 97.5% (mean = 66.7%) in the metalimnion. Finally, estimated viral mediated mortality rates from the percentage of visibly infected cells and potential nanoflagellate grazing rates based on assumed clearance rates suggest that flagellates consumed a larger proportion of bacterial production than was lost to viral lysis.     

Significance of Viral Lysis and Flagellate Grazing as Factors Controlling Bacterioplankton Production in a Eutrophic Lake

The effects of viral lysis and heterotrophic nanoflagellate (HNF) grazing on bacterial mortality were estimated in a eutrophic lake (Lake Plußsee in northern Germany) which was separated by a steep temperature and oxygen gradient into a warm and oxic epilimnion and a cold and anoxic hypolimnion. Two transmission electron microscopy-based methods (whole-cell examination and thin sections) were used to determine the frequency of visibly infected cells, and a model was used to estimate bacterial mortality due to viral lysis. Examination of thin sections also showed that between 20.2 and 29.2% (average, 26.1%) of the bacterial cells were empty (ghosts) and thus could not contribute to viral production. The most important finding was that the mechanism for regulating bacterial production shifted with depth from grazing control in the epilimnion to control due to viral lysis in the hypolimnion. We estimated that in the epilimnion viral lysis accounted on average for 8.4 to 41.8% of the summed mortality (calculated by determining the sum of the mortalities due to lysis and grazing), compared to 51.3 to 91.0% of the summed mortality in the metalimninon and 88.5 to 94.2% of the summed mortality in the hypolimnion. Estimates of summed mortality values indicated that bacterial production was controlled completely or almost completely in the epilimnion (summed mortality, 66.6 to 128.5%) and the hypolimnion (summed mortality, 43.4 to 103.3%), whereas in the metalimnion viral lysis and HNF grazing were not sufficient to control bacterial production (summed mortality, 22.4 to 56.7%). The estimated contribution of organic matter released by viral lysis of cells into the pool of dissolved organic matter (DOM) was low; however, since cell lysis products are very likely labile compared to the bulk DOM, they might stimulate bacterial production. The high mortality of bacterioplankton due to viral lysis in anoxic water indicates that a significant portion of bacterial production in the metalimnion and hypolimnion is cycled in the bacterium-virus-DOM loop. This finding has major implications for the fate and cycling of organic nutrients in lakes.     

Changes in Bacterial Community Composition and Dynamics and Viral Mortality Rates Associated with Enhanced Flagellate Grazing in a Mesoeutrophic Reservoir

Bacterioplankton from a meso-eutrophic dam reservoir was size fractionated to reduce (<0.8-μm treatment) or enhance (<5-μm treatment) protistan grazing and then incubated in situ for 96 h in dialysis bags. Time course samples were taken from the bags and the reservoir to estimate bacterial abundance, mean cell volume, production, protistan grazing, viral abundance, and frequency of visibly infected cells. Shifts in bacterial community composition (BCC) were examined by denaturing gradient gel electrophoresis (DGGE), cloning and sequencing of 16S rDNA genes from the different treatments, and fluorescence in situ hybridization (FISH) with previously employed and newly designed oligonucleotide probes. Changes in bacterioplankton characteristics were clearly linked to changes in mortality rates. In the reservoir, where bacterial production about equaled protist grazing and viral mortality, community characteristics were nearly invariant. In the “grazer-free” (0.8-μm-filtered) treatment, subject only to a relatively low mortality rate (~17% day⁻¹) from viral lysis, bacteria increased markedly in concentration. While the mean bacterial cell volume was invariant, DGGE indicated a shift in BCC and FISH revealed an increase in the proportion of one lineage within the beta proteobacteria. In the grazing-enhanced treatment (5-μm filtrate), grazing mortality was ~200% and viral lysis resulted in mortality of 30% of daily production. Cell concentrations declined, and grazing-resistant flocs and filaments eventually dominated the biomass, together accounting for >80% of the total bacteria by the end of the experiment. Once again, BCC changed strongly and a significant fraction of the large filaments was detected using a FISH probe targeted to members of the Flectobacillus lineage. Shifts of BCC were also reflected in DGGE patterns and in the increases in the relative importance of both beta proteobacteria and members of the Cytophaga-Flavobacterium cluster, which consistently formed different parts of the bacterial flocs. Viral concentrations and frequencies of infected cells were highly significantly correlated with grazing rates, suggesting that protistan grazing may stimulate viral activity.     

Seasonal Depth-Related Gradients in Virioplankton: Standing Stock and Relationships with Microbial Communities in Lake Pavin (France)

This study presents a depth-related survey of virioplankton abundance in Lake Pavin (Massif Central, France), in relation to the abundances of heterotrophic prokaryotes, picocyanobacteria (Pcy), autotrophic picoeukaryotes (Peu), and of autotrophic (ANF) and heterotrophic (HNF) nanoflagellates. The sampling strategy was designed to be representative of the physico-chemical gradients of the whole water column of the lake, and the seasonal variability as well. In mixolimnic surface waters, all communities were present and viral abundance peaked in summer and autumn. Viral abundance was significantly correlated (p < 0.001) with Pcy, Peu, and ANF, indicating that cyanophages and perhaps other phytoplankton viruses represent a significant pool of viral standing stocks in the mixolimnion of Lake Pavin. Microautotrophs were absent in the deep monimolimnic water masses, where viruses and heterotrophic prokaryotes exhibited highest seasonal abundances in summer and/or autumn and were significantly correlated (p < 0.001) to each other. This indicates that the anoxic monimolimnion of Lake Pavin is an exclusive habitat for viruses and heterotrophic prokaryotes. We conclude that in this habitat, host availability is prevalent over other factors (temperature, oxygen, nutrients, grazers) in favoring viral proliferation.     

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.     

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.     

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.     

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.     

Genetic Variation of the Bloom-Forming Cyanobacterium Microcystis aeruginosa within and among Lakes: Implications for Harmful Algal Blooms

To measure genetic variation within and among populations of the bloom-forming cyanobacterium Microcystis aeruginosa, we surveyed a suite of lakes in the southern peninsula of Michigan that vary in productivity (total phosphorus concentrations of ~10 to 100 μg liter⁻¹). Survival of M. aeruginosa isolates from lakes was relatively low (i.e., mean of 7% and maximum of 30%) and positively related to lake total phosphorus concentration (P = 0.014, r² = 0.407, n = 14). In another study (D. F. Raikow, O. Sarnelle, A. E. Wilson, and S. K. Hamilton, Limnol. Oceanogr. 49:482-487, 2004), survival rates of M. aeruginosa isolates collected from an oligotrophic lake (total phosphorus of ~10 μg liter⁻¹ and dissolved inorganic nitrogen:total phosphorus ratio of 12.75) differed among five different medium types (G test, P of <0.001), with higher survival (P = 0.003) in low-nutrient media (28 to 37% survival) than in high-nutrient media. Even with the relatively low isolate survivorship that could select against detecting the full range of genetic variation, populations of M. aeruginosa were genetically diverse within and among lakes (by analysis of molecular variance, Φ_sc = 0.412 [Φ_sc is an F-statistic derivative which evaluates the correlation of haplotypic diversity within populations relative to the haplotypic diversity among all sampled populations], P = 0.001), with most clones being distantly related to clones collected from lakes directly attached to Lake Michigan (a Laurentian Great Lake) and culture collection strains collected from Canada, Scotland, and South Africa. Ninety-one percent of the 53 genetically unique M. aeruginosa clones contained the microcystin toxin gene (mcyA). Genotypes with the toxin gene were found in all lakes, while four lakes harbored both genotypes possessing and genotypes lacking the toxin gene.     

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.     

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.     

Dissimilatory Arsenate and Sulfate Reduction in Sediments of Two Hypersaline, Arsenic-Rich Soda Lakes: Mono and Searles Lakes, California

A radioisotope method was devised to study bacterial respiratory reduction of arsenate in sediments. The following two arsenic-rich soda lakes in California were chosen for comparison on the basis of their different salinities: Mono Lake (~90 g/liter) and Searles Lake (~340 g/liter). Profiles of arsenate reduction and sulfate reduction were constructed for both lakes. Reduction of [⁷³As]arsenate occurred at all depth intervals in the cores from Mono Lake (rate constant [k] = 0.103 to 0.04 h⁻¹) and Searles Lake (k = 0.012 to 0.002 h⁻¹), and the highest activities occurred in the top sections of each core. In contrast, [³⁵S]sulfate reduction was measurable in Mono Lake (k = 7.6 ×10⁴ to 3.2 × 10⁻⁶ h⁻¹) but not in Searles Lake. Sediment DNA was extracted, PCR amplified, and separated by denaturing gradient gel electrophoresis (DGGE) to obtain phylogenetic markers (i.e., 16S rRNA genes) and a partial functional gene for dissimilatory arsenate reduction (arrA). The amplified arrA gene product showed a similar trend in both lakes; the signal was strongest in surface sediments and decreased to undetectable levels deeper in the sediments. More arrA gene signal was observed in Mono Lake and was detectable at a greater depth, despite the higher arsenate reduction activity observed in Searles Lake. A partial sequence (about 900 bp) was obtained for a clone (SLAS-3) that matched the dominant DGGE band found in deeper parts of the Searles Lake sample (below 3 cm), and this clone was found to be closely related to SLAS-1, a novel extremophilic arsenate respirer previously cultivated from Searles Lake.     

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.     

Significance of viral lysis and flagellate grazing as factors controlling bacterioplankton production in a eutrophic lake

The effects of viral lysis and heterotrophic nanoflagellate (HNF) grazing on bacterial mortality were estimated in a eutrophic lake (Lake Plusssee in northern Germany) which was separated by a steep temperature and oxygen gradient into a warm and oxic epilimnion and a cold and anoxic hypolimnion. Two transmission electron microscopy-based methods (whole-cell examination and thin sections) were used to determine the frequency of visibly infected cells, and a model was used to estimate bacterial mortality due to viral lysis. Examination of thin sections also showed that between 20.2 and 29.2% (average, 26.1%) of the bacterial cells were empty (ghosts) and thus could not contribute to viral production. The most important finding was that the mechanism for regulating bacterial production shifted with depth from grazing control in the epilimnion to control due to viral lysis in the hypolimnion. We estimated that in the epilimnion viral lysis accounted on average for 8.4 to 41.8% of the summed mortality (calculated by determining the sum of the mortalities due to lysis and grazing), compared to 51.3 to 91.0% of the summed mortality in the metalimninon and 88.5 to 94.2% of the summed mortality in the hypolimnion. Estimates of summed mortality values indicated that bacterial production was controlled completely or almost completely in the epilimnion (summed mortality, 66.6 to 128.5%) and the hypolimnion (summed mortality, 43.4 to 103.3%), whereas in the metalimnion viral lysis and HNF grazing were not sufficient to control bacterial production (summed mortality, 22.4 to 56.7%). The estimated contribution of organic matter released by viral lysis of cells into the pool of dissolved organic matter (DOM) was low; however, since cell lysis products are very likely labile compared to the bulk DOM, they might stimulate bacterial production. The high mortality of bacterioplankton due to viral lysis in anoxic water indicates that a significant portion of bacterial production in the metalimnion and hypolimnion is cycled in the bacterium-virus-DOM loop. This finding has major implications for the fate and cycling of organic nutrients in lakes.     

Decomposition Studies in Two Central Ontario Lakes Having Surficial pHs of 4.6 and 6.6

The rates of cellulose breakdown, composition of detrital microflora, and density of bacterial populations were determined in the epilimnetic sediments and water columns of two poorly buffered, oligotrophic, Canadian Shield lakes having mean surficial pHs of 4.6 (Bat Lake) and 6.6 (Harp Lake). The decomposition rate was significantly lower in oxic sediment of the acidified lake than of the circumneutral lake, but water column rates were almost identical in the two lakes. These results are explained in terms of the groups of cellulolytic microorganisms which were observed by phase-contrast microscopy as being active at the different sites: fungi in Bat Lake water and Cytophaga-like bacteria in the water and sediment of Harp Lake. Cytophaga-like bacteria were also the main decomposers in Bat Lake sediment, but their activity was restricted at porewater pHs of <5.0. Acridine orange direct counts of bacteria in the top centimeter of sediment ranged from 3.7 × 10⁸ to 1.0 × 10⁹ per g, and counts in planktonic water samples ranged from 4.9 × 10⁵ to 1.2 × 10⁶ per ml. Bacterial densities at most sites decreased significantly (P < 0.001) from August to late October, but did not show a consistent pattern of differences related to pH.