Marisol Felip

Lake Redó ecosystem response to an increasing warming in the Pyrennees during the twentieth century.

The ecosystem response of Lake Redó (Central Pyrenees) to fluctuations in seasonal air temperature during the last two centuries was investigated by comparison of reconstructed air temperatures with the sediment record. Fine slicing allowed a resolution of 3–6 years according to the 210Pb dating, although it was still difficult to easily investigate the response to air temperature forcing, since extreme fluctuations in temperature occur on interannual time-scales. However, the resolution was sufficient to show responses on decadal and century scales. An overall tendency to warming in mean annual temperature in the Central Pyrenees has been caused by summer and in particular by autumn increases. Many of the measured sediment variables apparently responded to these long term trends, but the significance of the relationships was highly conditioned by the structure of the data. The variables responding most on the finer time scales were the microfossils. For diatoms, chironomids and chrysophytes the main variability correlated to summer and to autumn temperatures. For two planktonic species, Fragilaria nanana and Cyclotella pseudostelligera, we found a link of their variability with temperature fluctuations in their growing months (September and October, respectively). This relationship appeared at a certain point during a general warming trend, indicating a threshold in the response. On the other hand, no significant changes in the dominant species could be linked to temperature, nor in any significant subgroup of the 180 diatom species present in the core. In contrast, for most chironomids (particularly Paratanytarsus austriacus, Heterotrissocladius marcidus and Micropsectra radialis) a negative relationship with summer temperature extended throughout the studied period. This response of the whole group gives chironomids a more robust role as indicators for recording temperature changes on long time-scales (e.g., through the Holocene) and for lake signal inter-comparison. Finally, our results indicated that, in all cases, there was a significant resilience to high frequency changes and hysteresis despite extreme fluctuations. Although we were dealing with organisms with one or many generations per year, their populations seemed to follow the decadal trends in air temperature.

Catalyzed Reported Deposition-Fluorescence In Situ Hybridization Protocol To Evaluate Phagotrophy in Mixotrophic Protists

ABSTRACT We describe a catalyzed reported deposition-fluorescence in situ hybridization (CARD-FISH) protocol particularly suited to assess the phagotrophy of mixotrophic protists on prokaryotes, since it maintains cell and plastid integrity, avoids cell loss and egestion of prey, and allows visualization of labeled prey against plastid autofluorescence. This protocol, which includes steps such as Lugols-formaldehyde-thiosulfate fixation, agarose cell attachment, cell wall permeabilization with lysozyme plus achromopeptidase, and signal amplification with Alexa-Fluor 488, allowed us to detect almost 100% of planktonic prokaryotes (Bacteria and Archaea) and, for the first time, to show archaeal cells ingested by mixotrophic protists.

Nitrogen in the Pyrenean lakes (Spain)

Lakes in the Pyrenees show a broad variability in nitrogen content and in the distribution of its different oxidation forms, which has no direct relation with any single physiographical, chemical or trophic feature of the lakes. Concentration of bound nitrogen in rain is low compared with other European mountains, but the annual load lies in the middle range. Seasonal and local variation in the composition of rainwater mainly depends on the geographical origin of the storms. Catchment and in-lake processes introduce further variability: NH4+, which is at similar concentration to NO3- in the rain, is quickly oxidized or adsorbed in the catchment; aquatic macrophytes can either reduce mean NO3- concentration in lake water (Ranunculo-Potametum) or greatly increase it in sediment pore water (Isoetes); NO2- depends on pH; decomposition of particulate nitrogen in sediments changes with depth; lakes act as traps for dissolved inorganic nitrogen; changes in dissolved organic nitrogen suggest high microbial activities even in cold waters; melting period introduces most of the seasonal variability.

Lake ice microbial communities in alpine and antarctic lakes

The observation that metabolic processes are reduced or completely inhibited at freezing temperatures has influenced microbial ecologists and limnologists, who have commonly concentrated on warm seasons and temperate habitats. High altitude and high latitude sites are not easily accessible and conducting research at low temperatures is not a trivial matter. However, methods for in-situ studies of microbial processes such as bacterial growth and production have been improved to a degree that allows the study of very oligotrophic systems at low temperatures. Furthermore, the construction of observatories and field stations has increased accessibility to high alpine and antarctic ecosystems, which has largely extended our knowledge of microbial life in cold environments.

Microbial food web components, bulk metabolism, and single-cell physiology of piconeuston in surface microlayers of high-altitude lakes

Sharp boundaries in the physical environment are usually associated with abrupt shifts in organism abundance, activity, and diversity. Aquatic surface microlayers (SML) form a steep gradient between two contrasted environments, the atmosphere and surface waters, where they regulate the gas exchange between both environments. They usually harbor an abundant and active microbial life: the neuston. Few ecosystems are subjected to such a high UVR regime as high altitude lakes during summer. Here, we measured bulk estimates of heterotrophic activity, community structure and single-cell physiological properties by flow cytometry in 19 high-altitude remote Pyrenean lakes and compared the biological processes in the SML with those in the underlying surface waters. Phototrophic picoplankton (PPP) populations, were generally present in high abundances and in those lakes containing PPP populations with phycoerythrin (PE), total PPP abundance was higher at the SML. Heterotrophic nanoflagellates (HNF) were also more abundant in the SML. Bacteria in the SML had lower leucine incorporation rates, lower percentages of “live” cells, and higher numbers of highly-respiring cells, likely resulting in a lower growth efficiency. No simple and direct linear relationships could be found between microbial abundances or activities and environmental variables, but factor analysis revealed that, despite their physical proximity, microbial life in SML and underlying waters was governed by different and independent processes. Overall, we demonstrate that piconeuston in high altitude lakes has specific features different from those of the picoplankton, and that they are highly affected by potential stressful environmental factors, such as high UVR radiation.

A comparative study of fluorescence-labelled enzyme activity methods for assaying phosphatase activity in phytoplankton. A possible bias in the enzymatic pathway estimations

We compared different fluorescence-labelled enzyme activity (FLEA) methods for assaying phosphatase activity in phytoplankton. Unfixed and liquid incubations are devised. We demonstrated that the presence of intracellular labelling was persistent, which could point out a source of bias in ectoenzymatic activities measurements based either on the FLEA or classical methods.

Some Mixotrophic Flagellate Species Selectively Graze on Archaea.

ABSTRACT Many phototrophic flagellates ingest prokaryotes. This mixotrophic trait becomes a critical aspect of the microbial loop in planktonic food webs because of the typical high abundance of these flagellates. Our knowledge of their selective feeding upon different groups of prokaryotes, particularly under field conditions, is still quite limited. In this study, we investigated the feeding behavior of three species (Rhodomonas sp., Cryptomonas ovata, and Dinobryon cylindricum) via their food vacuole content in field populations of a high mountain lake. We used the catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH) protocol with probes specific for the domain Archaea and three groups of Eubacteria: Betaproteobacteria, Actinobacteria, and Cytophaga-Flavobacteria of Bacteroidetes. Our results provide field evidence that contrasting selective feeding exists between coexisting mixotrophic flagellates under the same environmental conditions and that some prokaryotic groups may be preferentially impacted by phagotrophic pressure in aquatic microbial food webs. In our study, Archaea were the preferred prey, chiefly in the case of Rhodomonas sp., which rarely fed on any other prokaryotic group. In general, prey selection did not relate to prey size among the grazed groups. However, Actinobacteria, which were clearly avoided, mostly showed a size of <0.5 μm, markedly smaller than cells from the other groups. IMPORTANCE That mixotrophic flagellates are not randomly feeding in the main prokaryotic groups under field conditions is a pioneer finding in species-specific behavior that paves the way for future studies according to this new paradigm. The particular case that Archaea were preferentially affected in the situation studied shows that phagotrophic pressure cannot be disregarded when considering the distribution of this group in freshwater oligotrophic systems.

Experimental evidence of the quantitative relationship between the prokaryote ingestion rate and the food vacuole content in mixotrophic phytoflagellates: Mixotroph ingestion rates and vacuole content

The verification that many phytoflagellates ingest prokaryotes has changed the view of the microbial loop in aquatic ecosystems. Still, progress is limited because the phagotrophic activity is difficult to quantify in natural assemblages. Linking the food vacuole content in protist with the ingestion rate of prokaryotes would provide a crucial step forward. In this study, using the catalysed reporter deposition - fluorescence in situ hybridization protocol (CARD-FISH), which allows the visualization of labelled prokaryotes inside protists without relying on incubation procedures, we experimentally relate the food vacuole content of prokaryotes (Vc ) to the population-averaged ingestion rates (Ir ) estimated using bacteria-size fluorescent microspheres. The two variables relate according to the equation Ir = 7.52 Vc 0.9 , which indicates a prokaryote half-life of about 6 min in the protist vacuole. Five mixotrophic flagellate species from natural and culture populations were evaluated seven times during 24 h; they provided a broad range of average vacuole content (0.01 to 2.02 prokaryote protist-1 ) and ingestion rates (0.18 to 23 prokaryote protist-1 h-1 ). Consequently, the relationship found can be applied to quantify the mixotrophy activity in a large variety of field and experimental studies.