Anaerobic Microbial Communities in Lake Pavin, a Unique Meromictic Lake in France

The Bacteria and Archaea from the meromictic Lake Pavin were analyzed in samples collected along a vertical profile in the anoxic monimolimnion and were compared to those in samples from the oxic mixolimnion. Nine targeted 16S rRNA oligonucleotide probes were used to assess the distribution of Bacteria and Archaea and to investigate the in situ occurrence of sulfate-reducing bacteria and methane-producing Archaea involved in the terminal steps of the anaerobic degradation of organic material. The diversity of the complex microbial communities was assessed from the 16S rRNA polymorphisms present in terminal restriction fragment (TRF) depth patterns. The densities of the microbial community increased in the anoxic layer, and Archaea detected with probe ARCH915 represented the largest microbial group in the water column, with a mean Archaea/Eubacteria ratio of 1.5. Terminal restriction fragment length polymorphism (TRFLP) analysis revealed an elevated archaeal and bacterial phylotype richness in anoxic bottom-water samples. The structure of the Archaea community remained rather homogeneous, while TRFLP patterns for the eubacterial community revealed a heterogeneous distribution of eubacterial TRFs.     

Phylogenetic Diversity of Archaea and Bacteria in the Anoxic Zone of a Meromictic Lake (Lake Pavin, France)

The compositions of archaeal and bacterial populations at different depths (60 m [mixolimnion-chemocline interface], 70 m [chemocline-subchemocline interface], 90 m, and 92 m [the water-sediment interface]) in the anoxic zone of the water column in Lake Pavin, a freshwater permanently stratified mountain lake in France, were determined. Phylogenetic trees were constructed from sequences to assess archaeal and bacterial diversity at the four sites.     

Anaerobic microbial communities in Lake Pavin, a unique meromictic lake in France

The Bacteria and Archaea from the meromictic Lake Pavin were analyzed in samples collected along a vertical profile in the anoxic monimolimnion and were compared to those in samples from the oxic mixolimnion. Nine targeted 16S rRNA oligonucleotide probes were used to assess the distribution of Bacteria and Archaea and to investigate the in situ occurrence of sulfate-reducing bacteria and methane-producing Archaea involved in the terminal steps of the anaerobic degradation of organic material. The diversity of the complex microbial communities was assessed from the 16S rRNA polymorphisms present in terminal restriction fragment (TRF) depth patterns. The densities of the microbial community increased in the anoxic layer, and Archaea detected with probe ARCH915 represented the largest microbial group in the water column, with a mean Archaea/Eubacteria ratio of 1.5. Terminal restriction fragment length polymorphism (TRFLP) analysis revealed an elevated archaeal and bacterial phylotype richness in anoxic bottom-water samples. The structure of the Archaea community remained rather homogeneous, while TRFLP patterns for the eubacterial community revealed a heterogeneous distribution of eubacterial TRFs.     

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.     

Phylogenetic diversity of archaea and bacteria in the anoxic zone of a meromictic lake (Lake Pavin, France)

The compositions of archaeal and bacterial populations at different depths (60 m [mixolimnion-chemocline interface], 70 m [chemocline-subchemocline interface], 90 m, and 92 m [the water-sediment interface]) in the anoxic zone of the water column in Lake Pavin, a freshwater permanently stratified mountain lake in France, were determined. Phylogenetic trees were constructed from sequences to assess archaeal and bacterial diversity at the four sites.     

Seasonal changes in alkaline phosphatase activity of bacteria and microalgae in Lake Pavin (Massif Central,France)

Seasonal changes in alkalinephosphatase activity of bacteria and microalgae werestudied in the dimictic Lake Pavin (Massif Central,France), to test whetherthis activity is primarily algal or bacterial andwhether the APA presents seasonal variations coupledwith abiotic and biotic variables. Samples werecollected at different depths from May 1992 to May1993. The specific phosphatase activities wereanalysed spectrophotometrically with p-NitrophenylPhosphate (p-NPP) as substrate and were related to theprotein concentrations. No correlation was foundbetween alkaline phosphatase activity and solublereactive phosphorus (SRP) concentrations across anannual cycle. The specific activities of the smallclass (0.2–1.2 m) were the highest and thecontribution of this picoplanktonic size class(0.2–1.2 m) increased with depth. In addition, thelinear correlations between alkaline phosphataseactivity and protein concentration seemed to indicatethat most of these enzymes are constitutive. However,it cannot be excluded that the high phosphorusconcentrations repress APA.Finally, the measure of APA does not seem to be avalid quantitative test of the deficiency ofphosphorus for aquatic microorganisms.     

Reproduction,condition and food of adult arctic charr (Salvelinus alpinus,L.) in Lake Pavin (Massif Central,France)

RésuméTo determine the role and the importance of adult fish predation directly and indirectly in the food web of oligomesotrophic Lake Pavin, reproduction, condition, and feeding activity of the dominant adult chctic charr (Salvelinus alpinus, L.) were studied. Fish were caught by gillnets of different mesh sizes once a month from April to December 1992. The gonad cycle showed that the spawning period occurred between October to December. Relative fecundity is about 2509 ovules per kg of female fish and ovule mean diameter is 4.7 mm ( 0.6 mm). Seasonal variations of the sex-ratio (Max. in April = 2.67 and Min. in October=0.19) indicated the intralake migrations of females and males. The condition coefficients showed higher values for females than males; the maximum was observed in summer for both sexes and the minimum in April and in October for males and females, respectively. Low values of the condition were also observed during the spawning period. High feeding activity was recorded in spring and in December after reproduction; low feeding activity was observed in October during reproduction. Asellus aquaticus was quantitatively the dominant prey from April to September; during the sp awning period, S. alpinus consumed its own eggs, particularly in large quantity in December. Daphnia longispina was numerically the dominant prey all over the studied cycle and the electivity (Ivlev Index) of arctic charr on the latter prey ranged from +s+0.79 to +0.16. During April, more than 15 500 individuals of D. longispina per kg of fish were found in the gut contents. The selective predation of S. alpinus on benthic (A. aquatics) and zooplankton prey (D. longispina) indicated that the fish immigrate during their feeding activity from benthic/littoral to the pelagic area.Afin de d terminer le rôle et l'importance de la prédation directe et indirecte de l'omble chevalier adulte (Salvelinus alpinus, L.) dans le réseau trophique du lac Pavin, milieu oligo- mésotrophe, nous avons étudié la reproduction, deux facteurs de condition et l'activité alimentaire de ce poisson qui est l'espéce dominante dans ce lac. Les poissons ont été capturés par des filets maillants de différent vide de maille selon un pas d'échantillonnage mensuel d'avril á décembre 1992. Le cycle du développement des gonades montre que la période de reproduction se déroule entre octobre et décembre. La fécondité relative calculée est de 2509 ovules par kg de femelles et le diamétre moyen d'un ovule est de 4,7 mm ( 0,6). Les variations saisonniéres du sex-ratio (Max. en avril = 2,67 et Min. en octobre=0,19) indiquent le déplacement actif et successif des femelles et des mâles. Les variations saisonniéres des coefficients de condition indiquent une copulence plus importante des femelles par rapport aux mâles avec des valeurs maximales en été pour les deux sexes et des valeurs minimales en avril et en octobre pour les mâles et les femelles respectivement. De faibles valeurs des facteurs de condition ont été observées pendant la période de reproduction pour les deux sexes. L'activité alimentaire de l'omble chevalier est intense au printemps et au mois de décembre aprés la fraie et faible au mois d'octobre pendant la reproduction. Asellus aquaticus est, en termes de biomasse, la proie dominante d'avril á septembre. Pendant la période de reproduction, S. alpinus se nourrit de ses propres oeufs sur les zones de frayéres et en décembre la quantité d'oeufs consommée par le Salmonidé est importante. Daphnia longispina est, en termes d'abondance, la proie dominante tout au long de cette étude et l'indice d'électivité (Indice d'Ivlev) de l' ombre chevalier vis-á-vis de cette proie varie entre + 0,79 et + 0,16.De plus, au mois d'avril, plus de 15 500 D. longispina par kg de poisson ont été enregistrées dans les contenus stomacaux. La prédation sélective de S. alpinus sur des proies benthiques (A. aquaticus) et zooplanctoniques (D. longispina) indique que le poisson effectue des déplacements en fonction de son activité alimentaire de la zone littorale/benthique á la zone pélagique.     

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.     

Nitrogen Dynamics in the Steeply Stratified,Temperate Lake Verevi,Estonia

The dynamics of different nitrogen compounds and nitrification in diverse habitats of a stratified Lake Verevi (Estonia) was investigated in 2000–2001. Also planktonic N₂-fixation (N₂fix) was measured in August of the observed years. The nitrogen that accumulated in the hypolimnion was trapped in the non-mixed layer during most of the vegetation period causing a concentration of an order of magnitude higher than in the epilimnion. The ammonium level remained low in the epilimnion (maximum 577 mgN m⁻³, average 115 mgN m⁻³) in spite of high concentrations in the hypolimnion (maximum 12223 mgN m⁻³, average 4807 mgN m⁻³). The concentrations of NO₂⁻ and NO₃⁻ remained on a low level both in the epilimnion (average 0.94 and 9.09 mgN m⁻³, respectively) and hypolimnion (average 0.47 and 5.05 mgN m⁻³, respectively). N₂fix and nitrification ranged from 0.30 to 2.80 mgN m⁻³ day⁻¹ and 6.0 to 107 mgN m⁻³ day⁻¹, respectively; the most intensive processes occurred in 07.08.00 at depths of 2 and 5 m, accordingly. The role of N₂fix in the total nitrogen budget of Lake Verevi (in August 2000 and 2001) was negligible while episodically in the nitrogen-depleted epilimnion the N₂fix could substantially contribute to the pool of mineral nitrogen. Nitrification was unable to influence nitrogen dynamics in the epilimnion while some temporary coupling with ammonium dynamics in the hypolimnion was documented.     

Seasonal Depth-Related Gradients in Virioplankton: Lytic Activity and Comparison with Protistan Grazing Potential in Lake Pavin (France)

This study presents an original depth-related survey of virioplankton lytic activity in relation to prokaryotic production and potential protistan bacterivory in the deep (Z(max) = 92 m) meromictic volcanic Lake Pavin (Massif Central, France). The sampling strategy was designed to be representative of the physico-chemical gradients of the water column of the lake, and of the seasonal variability as well, i.e. 12 different depths sampled in triplicates from April to December 2005. In the space, viral lytic activity estimated from the frequency of visibly infected prokaryotic cells and from burst size over the study period generally decreased with depth. This was viewed as a paradox compared to the abundances of viruses and prokaryotes and to the prokaryotic production which increased with depth. The seasonal variability in viral lytic activity was correlated with prokaryotic variables (abundance and production) in the deepest waters, i.e. from the hypolimnion downwards. Compared to previous studies known from the mixolimnion, we conclude that the deep waters in Lake Pavin represent an exclusive environment for heterotrophic prokaryotes whose seasonal activity offers an optimal and unique resource for thriving viral communities, some of which may be typical, endemic to the ambient dark, cold and stable deep water masses. Overall, the main findings in the present study get well around a previous statement that the ecology of the deepest waters of Lake Pavin is essentially driven by the dark viral loop (dissolved organic matter-prokaryotes-viruses) processes, which can sequester organic matters and nutrients for a long-lived turnover time. This is in agreement with recent demonstrations from marine systems that meso- and bathypelagic waters are optimal environments for viral survival and proliferation.     

Viral abundance, production, decay rates and life strategies (lysogeny versus lysis) in Lake Bourget (France)

We have investigated the ecology of viruses in Lake Bourget (France) from January to August 2008. Data were analysed for viral and bacterial abundance and production, viral decay, frequency of lysogenic cells, the contribution of bacteriophages to prokaryotic mortality and their potential influence on nutrient dynamics. Analyses and experiments were conducted on samples from the epilimnion (2 m) and the hypolimnion (50 m), taken at the reference site of the lake. The abundance of virus‐like particles (VLP) varied from 3.4 × 10⁷ to 8.2 × 10⁷ VLP ml^?1; with the highest numbers and virus‐to‐bacterium ratio (VBR = 69) recorded in winter. Viral production varied from 3.2 × 10⁴ VLP ml^?1 h^?1 (July) to 2 × 10⁶ VLP ml^?1 h^?1 (February and April), and production was lower in the hypolimnion. Viral decay rate reached 0.12–0.15 day^?1, and this parameter varied greatly with sampling date and methodology (i.e. KCN versus filtration). Using transmission electron microscopy (TEM) analysis, viral lysis was responsible for 0% (January) to 71% (February) of bacterial mortality, while viral lysis varied between 0% (April) and 53% (January) per day when using a modified dilution approach. Calculated from viral production and burst size, the virus‐induced bacterial mortality varied between 0% (January) and 68% (August). A weak relationship was found between the two first methods (TEM versus dilution approach). Interestingly, flow cytometry analysis performed on the dilution experiment samples revealed that the viral impact was mostly on high DNA content bacterial cells whereas grazing, varying between 8.3% (June) and 75.4% (April), was reflected in both HDNA and LDNA cells equally. The lysogenic fraction varied between 0% (spring/summer) and 62% (winter) of total bacterial abundance, and increased slightly with increasing amounts of mitomycin C added. High percentages of lysogenic cells were recorded when bacterial abundance and activity were the lowest. The calculated release of carbon and phosphorus from viral lysis reached up to 56.5 µgC l^?1 day^?1 (assuming 20 fgC cell^?1) and 1.4 µgP l^?1 day^?1 (assuming 0.5 fgP cell^?1), respectively, which may represent a significant fraction of bacterioplankton nutrient demand. This study provides new evidence of the quantitative and functional importance of the virioplankton in the functioning of microbial food webs in peri‐alpine lakes. It also highlights methodologically dependent results.     

Photolithotrophy,photoheterotrophy and chemoheterotrophy during spring phytoplankton development (Lake Pavin)

In order to determine the relative importance of autotrophic and heterotrophic activities in both bacterial and phytoplanktonic communities in an oligomesotrophic lake, the size fractionation by differential filtration and the use of a bacterial inhibitor (gentamycin) were combined. The study was carried out at Lake Pavin during the spring planktonic bloom. Photosynthetic and photo- and chemoheterotrophic activities were measured from the assimilation of NaH¹⁴CO₃ and glucose-³H, using a double labeling technique. The bacterial community was at low cell concentration (0.6 to 7 × 10⁵ cells ml^–) and represented very low biomass values (0.9 to 11.5 gC liter^–1). The abundance of the phytoplankton varied between 0.5 and 1.8 × 10⁶ cells liter^–1, and biomass values ranged between 19 and 118 gC liter^–1. The diatom Melosira italica sp. subarctica (O. Mueller) was the largely dominant species in the meta- and hypolimnion. Inorganic fixation by photolithotrophy (mean value: 1.66 mg C m^–3 hour^–1) largely predominates over assimilation by photoheterotrophy (mean value: 0.93 g C m^–3 hour^–1) or chemoheterotrophy (mean value: 2.42 g C m^–3 hour^–1). However, because of the considerable underestimation of heterotrophic assimilation due to the experimental methods used, and because of the spatial and temporal separation of photolithotrophic and photo- and chemoheterotrophic activities, it is likely that the fixation of organic carbon by microalgae plays an important role in the survival of species and/or in competitive interactions, as the results with Monoraphidium contortum (Pasch. et Korschik.), the prevailing species in the epilimnion, would suggest. Send offprint requests to: C. Amblard.     

The physical structure and dynamics of a deep,meromictic crater lake (Lac Pavin,France)

The characteristic feature of the physical structure of Lac Pavin is a distinct and permanent chemically induced density increase between about 60 and 70 m depth. This chemocline separates the seasonally mixed mixolimnion from the monimolimnion, which is characterized by elevated temperature and salinity as well as complete anoxia. Previously published box-models of the lake postulated substantial groundwater input at the lake bottom, and consequently a short water residence time in the monimolimnion and high fluxes of dissolved constituents across the chemocline. We present a new view of the physical structure and dynamics of Lac Pavin, which is based on the results of high-resolution CTD profiles, transient and geochemical tracers (tritium, CFCs, helium), and numerical modeling. The CTD profiles indicate the existence of a sublacustrine spring above rather than below the chemocline. A stability analysis of the water column suggests that vertical turbulent mixing in the chemocline is very weak. A numerical one-dimensional lake model is used to reconstruct the evolution of transient tracer distributions over the past 50 years. Model parameters such as vertical diffusivity and size of sublacustrine springs are constrained by comparison with observed tracer data. Whereas the presence of a significant water input to the monimolimnion can clearly be excluded, the input to the mixolimnion – both at the surface and from the indicated sublacustrine spring – cannot be accurately determined. The vertical turbulent diffusivity in the chemocline is well constrained to K 5×10^-8 m² s^-1, about a factor of three below the molecular diffusivity for heat. Assuming thus purely molecular heat transport, the heat flow through the chemocline can be estimated to between 30 and 40 mW m^-2. With respect to dissolved constituents, the very weak turbulent diffusive exchange is equivalent to a stagnant monimolimnion with a residence time of nearly 100 years. Based on these results and vertical concentration profiles of dissolved species, diffusive fluxes between monimolimnion and mixolimnion can be calculated. A large excess of helium with a ³He/⁴He ratio of (9.09 ± 0.01)×10^-6 (6.57 R _a) is present in the monimolimnion, clearly indicating a flux of magmatic gases into the monimolimnion. We calculate a flux of 1.0×10^-12 mol m^-2 s^-1 for mantle helium and infer a flux of 1.2×10^-7 mol m^-2 s^-1 (72 t year^-1) for magmatic CO₂. The monimolimnion appears to be in steady state with respect to these fluxes.     

Evidence for a diurnal variation in the carotenoid content of acanthodiaptomus denticornis (Crustacea,copepoda) in Lac Pavin (Auvergne,France)

In Lac Pavin Acanthodiaptomus denticornis was found to be intensely reddish-orange coloured by keto-carotenoids of the astaxanthin type. Such pigments are not normally found in phytoplankton algae, and apparently these carotenoids result from the metabolism of pigments of dietary origin. The carotenoid content of the zooplankton showed a distinct 2.5-fold diurnal variation, with a minimum at night time and a maximum in the early morning. The possible impact of a diurnal difference in zooplankton feeding activity is discussed.     

Evolution de la composition en acides gras d'un phytoplancton lacustre (Lac Pavin,France)

Fatty Acid Composition of Lacustrian Phytoplankton (Pavin Lake, France) Fatty acid composition of lacustrian phytoplankton was established during a study carried out on Lake Pavin (France) from May to September 1981. Fatty acid spectra point out even numbers of carbon atoms (ranging from 14 to 22) and in particular unsaturated fatty acids preponderance. In addition, the occurrence of some unsaturated fatty acids is directly linked to phytoplankton taxonomic composition, but also to physiological state of the cells. Lastly, the main types of phytoplankton populations differ in the degree of unsaturation of their total fatty acids.     

The Characteristics of a Temperate Eutrophic,Dimictic Lake (Lake Mikołajskie,Northern Poland)

Gross primary production equals 3160 kcal Xm⁻² ×yr⁻¹, 38% coming from the littoral. The efficiencies of both nonpredatory and predatory zooplankton production are high. The production of phytoplankton is expended mostly for sedimentation in April, for grazing by zooplankton in June, heterotrophic respiration and cumulation of dissolved and particulate organic matter in August—September and sedimentation in late autumn. Almost total elimination of zooplankton is done by predators, mostly invertebrates. About 10% of autochthonically produced plus allochthonic matter is annually removed from cycling, mostly as permanent bottom deposits. The annual load of phosphorus is 3 times higher than the permissible VOLLENWEIDER'S level. About 70% of phosphorus load, but only two percent of its reserve in the lake is annually removed from cycling. Progressing eutrophication resulted in several times increased phytoplankton biomass, decrease of phytoplankton P/B and share of nannoplankton, increase of decomposition in epilimnion, etc, during 10 years. Inspite of this, pelagic community in spring still has rather mesotrophic than highly eutrophic character. Increased eutrophication due to human impact (deforestation, agriculture, erosion) is also seen in bottom deposits since the 15th century.