Effects of (p)ppGpp on the progression of the cell cycle of Caulobacter crescentus.

Bacteria must control the progression of their cell cycle in response to nutrient availability. This regulation can be mediated by guanosine tetra- or pentaphosphate [(p)ppGpp], which are synthesized by enzymes of the RelA/SpoT homologue (Rsh) family, particularly under starvation conditions. Here, we study the effects of (p)ppGpp on the cell cycle of Caulobacter crescentus, an oligotrophic bacterium with a dimorphic life cycle. C. crescentus divides asymmetrically, producing a motile swarmer cell that cannot replicate its chromosome and a sessile stalked cell that is replication competent. The swarmer cell rapidly differentiates into a stalked cell in appropriate conditions. An artificial increase in the levels of (p)ppGpp in nonstarved C. crescentus cells was achieved by expressing a truncated relA gene from Escherichia coli, encoding a constitutively active (p)ppGpp synthetase. By combining single-cell microscopy, flow cytometry approaches, and swarming assays, we show that an increase in the intracellular concentration of (p)ppGpp is sufficient to slow down the swarmer-to-stalked cell differentiation process and to delay the initiation of chromosome replication. We also present evidence that the intracellular levels of two master regulators of the cell cycle of C. crescentus, DnaA and CtrA, are modulated in response to (p)ppGpp accumulation, even in the absence of actual starvation. CtrA proteolysis and DnaA synthesis seem indirectly inhibited by (p)ppGpp accumulation. By extending the life span of the motile nonreproductive swarmer cell and thus promoting dispersal and foraging functions over multiplication under starvation conditions, (p)ppGpp may play a central role in the ecological adaptation of C. crescentus to nutritional stresses.

Pollution stress on aquatic microbial communities

Thousands of chemical compounds enter the natural environment but many have unknown effects and consequences, in particular at low concentrations. This thesis work contributes to our understanding of pollution effects by using bacteria as test organisms. Bacteria are important for this question because some of them degrade and transform pollutants into less harmful compounds, but secondly because they themselves can be inhibited in their reproduction by exposure to toxic compounds. When inhibitory effects occur this may change the composition of the microbial com¬munity in the long run, leading to altered or diminished ecosystem services by those communities. As a result chemicals of anthropogenic origin may accumulate and per¬sist in the environment, and finally, affect higher organisms as well. In addition to acquiring basic understanding of pollutant effects at low concentrations on bacterial communities an applied goal of this thesis work was to develop bacteria-based tests to screen new organic chemicals for toxicity and biodégradation. In the first part of this work we developed a flow cytometry-based assay on SYT09 plus ethidium-bromide or propidium-iodide stained cells of Pseudomonas ûuorescens exposed or not to a variety of pollutants under oligotrophic growth conditions. Flow cytometry (FC) allows fast and accurate counting of bacterial cells under simul¬taneous assessment of their physiological state, in particular in combination with different fluorescent dyes. Here we employed FC and fluorescent dyes to monitor the effect that pollutants may exert on Pseudomonas ûuorescens SV3. First we designed an oligotrophic growth test, which enabled us to follow population growth at low densities (104 - 10 7 cells per ml) using 0.1 mM sodium acetate as carbon source. Cells in the oligotrophic milieu were then exposed or not to a variety of common pollutants, such as 2-chlorobiphenyl (2CBP), naphthalene (NAH), 4-chlorophenol (4CP), tetradecane (TD), mercury chloride (HgCl2) or benzene, in different dosages. Exposed culture samples were stained with SYT09 (green fluorescent dye binding nucleic acids, generally staining all cells) in combination with propidium iodide (PI) or ethidium bromide (EB), both dyes being membrane integrity indicators. We ob- served that most of the tested compounds decreased population growth in a dosage- dependent manner. SYT09/PI or SYT09/EB staining then revealed that chemical exposure led to arisal of subpopulations of live and injured or dead cells. By modeling population growth on the total cell numbers in population or only the subpopulation of live cells we inferred that even in stressed populations live cells multiply at rates no different to unexposed controls. The net decrease in population growth would thus be a consequence of more and more cells being not able to multiply at all, rather than all cells multiplying at slower rates. In addition, the proportion of injured cells correlated to the compound dosage. We concluded that the oligotrophic test may be useful to asses toxicity of unknown chemicals on a variety of model bacteria. Mul¬tiple tests can be run in parallel and effects are rapidly measured within a period of 8 hours. Interestingly, in the same exposure tests with P. fluorescens SV3 we observed that some chemicals which did not lead to a reduction of net population growth rates did cause measurable effects on live cells. This was mainly observed in cells within the live subpopulation as an increase of the EB fluorescence signal. We showed that SYT09/EB is a more useful combination of dyes than SYT09/PI because PI fluorescence tend to increase only when cells are effectively dead, but not so much in live cells (less then twofold). In contrast, EB geometric mean fluorescence in live cells increased up to eightfold after exposure to toxic compounds. All compounds even at the lowest concentration caused a measurable increase in EB geometric mean fluorescence especially after 2 h incubation time. This effect was found to be transient for cells exposed to 2CBP and 4CP, but chronic for cells incubated with TD and NAH (ultimately leading to cell death). In order to understand the mechanism underlying the observed effects we used known membrane or energy uncouplers. The pattern of EB signal increase in chemical-exposed populations resembled mostly that of EDTA, although EB fluorescence in EDTA-treated or pasteurized cells was even higher than after exposure to the four test chemicals. We conclude that the ability of cells to efflux EB under equilibrium conditions is an appropriate measure for the potential of a chemical to exert toxicity. Since most bacterial species possess efflux systems for EB that all require cellular energy, our test should be more widely relevant to infer toxicity effects of chemical exposure on the physiological status of the bacterial cell. To better understand the effect of toxicant exposure on efflux defense systems, we studied 2-hydroxybiphenyl toxicity to Pseudomonas azeiaica HBP1. We showed that 2-HBP exerts toxicity even to P. azelaica HBP1, but only at concentrations higher than 0.5 mM. Above this concentration transient loss of membrane polarization and integrity occurred, which we conclude from staining of growing cells with fluorescent dyes. Cells finally recover and resume growth on 2HBP. The high resistance of P. azelaica HBP1 to 2-HBP was found to be the result of an efficient MexABOprM- type efflux pump system counteracting passive influx of this compound into the membrane and cellular interior. Mutants with disrupted mexA, mexB and oprM genes did no longer grow on 2-HBP at concentrations above 100 μΜ, whereas below this concentration we found 2-HBP-concentration dependent decrease of growth rate. The MexAB-OprM system in P. azeiaica HBP1 is indeed an efflux pump for ethidium bromide as well. By introducing gfp reporter fusions responsive to intracellular 2- HBP concentrations into HBP1 wild-type or the mutants we demonstrated that 2HBP enters into the cells in a similar way. In contrast, the reporter system in the wild-type cells does not react to 2-HBP at an outside concentration of 2.4 μΜ, whereas in mutant cells it does. This suggests that wild-type cells pump 2-HBP to the outside very effectively preventing accumulation of 2-HBP. 2HBP metabolism, therefore, is not efficient enough to lower the intracellular concentration and prevent toxicity. We conclude that P. azelaica HBP1 resistance to 2-HBP is mainly due to an efficient efflux system and that 2HBP in high concentrations exerts narcotic effects on the bacterial membrane. In the part of this thesis, we investigated the possibilities of bacteria to degrade pollutants at low concentrations (1 mg per L and below). As test components we used 2-hydroxybiphenyl, antibiotics and a variety of fragrances, many of which are known to be difficult to biodegrade. By using accurate counting of low numbers of bacterial cells we could demonstrate that specific growth on these compounds is possible. We demonstrated the accuracy of FC counting at low cell numbers (down to 103 bacterial cells per ml). Then we tested whether bacterial population growth could be specifically monitored at the expense of low substrate concentrations, us¬ing P. azelaica HBP1. A perfect relationship was found between growth rate, yield and 2-HBP concentrations in the range of 0.1 up to 5 mg per L. Mixing P. azelaica within sludge, however, suggested that growth yields in a mixed community can be much lower than in pure culture, perhaps because of loss of metabolic intermediates. We then isolated new strains from activated sludge using 2-HBP or antibiotics (Nal, AMP, SMX) at low concentrations (0.1-1 mg per L) as sole carbon and energy sub¬strate and PAO microdishes. The purified strains were then examined for growth on their respective substrate, which interestingly, showed that all strains can not with¬stand higher than 1 or 10 mg per L concentrations of target substrate. Thus, bacteria must exist that contribute to compound degradation at low pollutant concentrations but are inhibited at higher concentrations. Finally we tested whether specific biomass growth (in number of cells) at the expense of pollutants can also be detected with communities as starting material. Hereto, we focused on a number of fragrance chemicals and measured community biomass increase by flow cytometry cell counting on two distinct starter communities: (i) diluted Lake Geneva water, and dilute activated sludge from a wastewater treatment plant. We observed that most of the test compounds indeed resulted in significant biomass increase in the starter community compared to a no-carbon added control, but activated sludge and lake Geneva water strongly differed (almost mutually ex¬clusive) in their capacity to degrade the test chemicals. In two cases for activated sludge the same type of microbial community developed upon compound exposure, as concluded from transcription fragment length polymorphism analysis on community purified and PCR amplified 16S rRNA gene fragments. To properly test compound biodegradability it is thus important to use starter communities of different origin. We conclude that FC counting can be a valuable tool to screen chemicals for their biodegradability and toxicity. - Des milliers de produits chimiques sont libérés dans l'environnement mais beaucoup ont des effets inconnus, en particulier à basses concentrations. Ce travail de thèse contribue à notre comprehension des effets de la pollution en utilisant des bacteries comme des organismes-tests. Les bacteries sont importantes pour etudier cette ques¬tion car certaines d'entre elles peuvent degrader ou transformer les polluants, mais également parce qu'elles-mmes peuvent tre inhibees dans leur reproduction après avoit ete exposees à ces composes toxiques. Quand des effets inhibiteurs ont lieu, la composition de la communauté microbienne peut tre changee à long terme, ce qui mène à une reduction du service d'ecosystème offert par ces communautés. En consequence, après leur liberation dans l'environnement, les produits chimiques d'origine anthropogenique peuvent soit s'y accumuler et per¬sister, exerant ainsi des effets encore inconnus sur les organismes vivants. En plus d'acquérir des connaissances de base sur les effets des polluants à basses concentra¬tions sur les communautés microbiennes, un but applique de cette thèse était de développer des tests bases sur les bacteries afin d'identifier de nouveau composes pour leur toxicité ou leur biodégradation. Dans la première partie de ce travail, nous avons developpe un test base sur la cytometrie de flux (FC) sur des cellules de Pseudomonas fluorescens colorees par du bromure d'ethidium ou de l'iodure de propidium et exposees ou non à une palette de polluants sous des conditions de croissance oligotrophique. La cytometrie de flux est une technique qui connaît de nombreuses applications dans la microbiologie environ¬nementale. Cela est principalement du au fait qu'elle permet un comptage rapide et precis ainsi que l'évaluation de l'état physiologique, en particulier lorsqu'elle est combinée h des colorations fluorescentes. Ici, nous avons utilise la technique FC et des colorants fluorescents afin de mesurer l'effet que peuvent exercer certains pollu¬ants sur Pseudomonas ûuorescens SV3 . D'abord nous avons conu des tests oligo- trophiques qui nous permettent de suivre la croissance complète de cellules en culture h des densites faibles (104 -10 7 cellules par ml), sur de l'acetate de sodium à 0.1 mM, en presence ou absence de produits chimiques (2-chlorobiphenyl (2CBP), naphthalène (NAH), 4-chlorophenol (4CP), tetradecane (TD), chlorure de mercure(II) (HgCl2)) à différentes concentrations. Afin de montrer le devenir des bacteries tant au niveau de la cellule individuelle que celui de la population globale, après exposition à des series de composes chimiques, nous avons compte les cellules colorees avec du SYT09 (col¬orant fluorescent vert des acides nucléiques pour la discrimination des cellules par rapport au bruit de fond) en combinaison avec l'iodure de propidium (PI) ou le bromure d'ethidium (EB), indicateurs de l'intégrité de la membrane cellulaire avec FC. Nous avons observe que de nombreux composes testes avaient un effet sur la croissance bacterienne, resultant en une baisse du taux de reproduction de la pop¬ulation. En outre, la double coloration que nous avons utilisee dans cette etude SYT09/PI ou SYT09/EB a montre que les produits chimiques testes induisaient une reponse heterogène des cellules dans la population, divisant celle-ci en sous- populations "saine", "endommagee" ou "morte". Les nombres de cellules à partir du comptage et de la proportion de celles "saines" et "endommagees/mortes" ont ensuite ete utilises pour modeliser la croissance de P. ûuorescens SV3 exposee aux produits chimiques. La reduction nette dans la croissance de population est une consequence du fait que de plus en plus de cellules sont incapables de se reproduire, plutt que du fait d'une croissance plus lente de l'ensemble de la population. De plus, la proportion de cellules endommagees est correllee au dosage du compose chimique. Les résultats obtenus nous ont permis de conclure que le test oligotrophique que nous avons developpe peut tre utilise pour l'évaluation de la toxicité de produits chimiques sur différents modèles bacteriens. Des tests multiples peuvent tre lances en parallèle et les effets sont mesures en l'espace de huit heures. Par ailleurs, nous en déduisons que les produits chimiques exercént un effet sur la croissance des cellules de P. ûuorescens SV3, qui est heterogène parmi les cellules dans la population et depend du produit chimique. Il est intéressant de noter que dans les mmes tests d'exposition avec P. ûuorescens SV3, nous avons observe que certains composes qui n'ont pas conduit à une reduction du taux de la croissance nette de la population, ont cause des effets mesurables sur les cellule saines. Ceci a ete essentiellement observe dans la portion "saine" des cellules en tant qu'augmentation du signal de la fluorescence de 1ΈΒ. D'abord nous avons montre que SYT09/EB était une com¬binaison de colorants plus utile que celle de SYT09/PI parce que la fluorescence du PI a tendance à augmenter uniquement lorsque les cellules sont effectivement mortes, et non pas dans les cellules saines (moins de deux fois plus). Par opposi¬tion, la fluorescence moyenne de l'EB dans les cellules saines augmente jusqu'à huit fois plus après exposition aux composes toxiques. Tous les composes, mme aux plus basses concentrations, induisent une augmentation mesurable de la fluorescence moy¬enne de 1ΈΒ, plus particulièrement après deux heures d'incubation. Cet effet s'est revele tre transitoire pour les cellules exposees aux 2CNP et 4CP, mais est chro¬nique pour les cellules incubees avec le TD et le NAH (entranant la mort cellulaire). Afin de comprendre les mécanismes qui sous-tendent les effets observes, nous avons utilise des decoupleurs d'energie ou de membrane. L'augmentation du signal EB dans les populations causee par des produits chimiques ressemblait à celle exerce par le chelateur des ions divalents EDTA. Cependant, les intensités du signal EB des cellules exposees aux produits chimiques testees n'ont jamais atteint les valeurs des cellules traitees avec l'EDTA ou pasteurises. Nous en concluons que le test oli- gotrophique utilisant la coloration (SYT09/)EB des cellules exposees ou non à un produit chimique est utile afin d'evaluer l'effet toxique exerce par les polluants sur la physiologie bacterienne. Afin de mieux comprendre la reaction d'un système de defense par pompe à efflux après exposition à une toxine, nous avons étudié la toxicité du 2-hydroxybiphenyl (2-HBP) sur Pseudomonas azeiaica HBP1. Nous avons montre que le 2-HBP exerce une toxicité mme sur HBP1, mais uniquement à des concentrations supérieures à 0.5 mM. Au-dessus de cette concentration, des pertes transitoires d'intégrité et de polarization membranaire ont lieu, comme cela nous a ete montre par coloration des cellules en croissance. Les cellules sont finalement capables de se rétablir et de reprendre leur croissance sur 2-HBP. La forte resistance de P. azeiaica HBP1 h 2-HBP physiologie bacterienne s'est revele tre le résultat d'un système de pompe h efflux de type MexABOprM qui contre-balance l'influx passif de ce compose h travers la membrane. Nous avons montre, en construisant des mutants avec des insertions dans les gènes mexA, mexB and oprM et des fusions avec le gène rapporteur gfp, que l'altération de n'importe quelle partie du système d'efflux conduisait à accroître l'accumulation de 2-HBP dans la cellule, en comparaison avec la souche sauvage HBP1, provoquant une diminution de la resistance au 2-HBP ainsi qu'une baisse du taux de reproduction des cellules. Des systèmes d'efflux similaires sont répandus chez de nombreuses espèces bactériennes. Ils seraient responsables de la resistance aux produits chimiques tels que les colorants fluorescents (bromure d'ethidium) et des antibiotiques. Nous concluons que la resistance de P. azelaica HBP1 à 2-HBP est principalement due à un système d'efflux efficace et que 2-HBP, à des concentrations elevees, exerce un effet deletère sur la membrane bacterienne. En se basant sur le comptage des cellules avec la FC, nous avons developpe ensuite une methode pour evaluer la biodegradabilite de polluants tels que le 2-HBP ainsi que les antibiotiques (acide nalidixique (Nal), ampicilline (AMP) ou sulfamethoxazole (SMX)) à de faibles concentrations lmg par L et moins), par le suivi de la croissance spécifique sur le compose de cultures microbiennes pures et mixtes. En utilisant un comptage precis de faibles quantités de cellules nous avons pu demontrer que la croissance spécifique sur ces composes est possible. Nous avons pu illustrer la precision du comptage par cytometrie de flux à faible quantité de cellules (jusqu'à 10 3 cellules par ml). Ensuite, nous avons teste s'il était possible de suivre dynamiquement la croissance de la population de cellules sur faibles concentrations de substrats, en utilisant P. azelaica HBP1. Une relation parfaite a ete trouvee entre le taux de croissance, le rendement et les concentrations de 2-HBP (entre 0.1 et 5 mg par L). En mélangeant HBP1 à de la boue active, nous avons pu montrer que le rendement en communauté mixtes pouvait tre bien inférieur qu'en culture pure. Ceci étant peut tre le résultat d'une perte d'intermédiaires métaboliques. Nous avons ensuite isole de nouvelles souches à partir de la boue active en utilisant le 2-HBP ou des antibiotiques (Nal, AMP, SMX) h basses concentrations (0.1-1 mg par L) comme seules sources de carbone et d'energie. En combinaison avec ceci, nous avons également utilise des microplaques PAO. Les souches purifiees ont ensuite ete examinees pour leurs croissances sur leurs substrats respectifs. De faon intéressante, toutes ces souches ont montre qu'elles ne pouvaient pas survivre à des concentrations de substrats supérieures à 1 ou 10 mg par L. Ainsi, il existe des bacteries qui contribuent à la degradation de composes à basses concentrations de polluant mais sont inhibes lorsque ces concentrations deviennent plus hautes. Finalement, nous avons cherche à savoir s'il est possible de detecter une croissance spécifique à une biomasse au depend d'un polluant, en partant d'une communauté microbienne. Ainsi, nous nous sommes concentre sur certains composes et avons mesure l'augmentation de la biomasse d'une communauté grce à la cytometrie de flux. Nous avons compte deux communautés de depart distinctes: (i) une dilution d'eau du Lac Léman, et une dilution de boue active d'une station d'épuration. Nous avons observe que la plupart des composes testes ont entrane une augmentation de la biomasse de depart par rapport au control sans addition de source de carbone. Néanmoins, les échantillons du lac Léman et de la station d'épuration différaient largement (s'excluant mutuellement l'un l'autre) dans leur capacité à degrader les composes chimiques. Dans deux cas provenant de la station d'épuration, le mme type de communauté microbienne s'est developpe après exposition aux composes, comme l'a démontré l'analyse TRFLP sur les fragments d'ARN 16S purifie de la communauté et amplifie par PCR. Afin de tester correctement la biodegradabilite d'un compose, il est donc important d'utiliser des communautés de depart de différentes origines Nous en concluons que le comptage par cytometrie de flux peut tre un outil de grande utilité pour mettre en valeur la biodegradabillite et la toxicité des composes chimiques.

Stratigraphic, sedimentological and palaeoenvironmental constraints on the rise of the Urgonian platform in the western Swiss Jura

Urgonian-type carbonates are a characteristic feature of many late Early Cretaceous shallow-marine, tropical and subtropical environments. The presence of typical photozoan carbonate-producing communities including corals and rudists indicates the prevalence of warm, transparent and presumably oligotrophic conditions in a period otherwise characterized by the high density of globally occurring anoxic episodes. Of particular interest, therefore, is the exploration of relationships between Urgonian platform growth and palaeoceanographic change. In the French and Swiss Jura Mountains, the onset and evolution of the Urgonian platform have been controversially dated, and a correlation with other, better dated, successions is correspondingly difficult. It is for this reason that the stratigraphy and sedimentology of a series of recently exposed sections (Eclepens, Vaumarcus and Neuchatel) and, in addition, the section of the Gorges de l'Areuse were analysed. Calcareous nannofossil biostratigraphy, the evolution of phosphorus contents of bulk rock, a sequence-stratigraphic interpretation and a correlation of drowning unconformities with better dated sections in the Helvetic Alps were used to constrain the age of the Urgonian platform. The sum of the data and field observations suggests the following evolution: during the Hauterivian, important outward and upward growth of a bioclastic and oolitic carbonate platform is documented in two sequences, separated by a phase of platform drowning during the late Early Hauterivian. Following these two phases of platform growth, a second drowning phase occurred during the latest Hauterivian and Early Barremian, which was accompanied by significant platform erosion and sediment reworking. The Late Barremian witnessed the renewed installation of a carbonate platform, which initiated with a phase of oolite production, and which progressively evolved into a typical Urgonian carbonate platform colonized by corals and rudists. This phase terminated at the latest in the middle Early Aptian, due to a further drowning event. The evolution of this particular platform segment is compatible with that of more distal and well-dated segments of the same northern Tethyan platform preserved in the Helvetic zone of the Alps and in the northern subalpine chains (Chartreuse and Vercors).

Impact of free-range pigs on mountain pastures in the Swiss Jura

The consequences of foraging by free-range pigs on the vegetation of mountain pastures was investigated. 25 pigs (15 week-old, mean weight 50 kg) were enclosed from June to mid-September in a 2 ha-enclosure in Jura Mountains (Switzerland), fed with a mixture of lactoserum and cereals. The enclosure contained five different plant communities. Eutrophic pastures on deep soil were strongly overturned, but the recolonisation was quick and dominated by the original species. Mesotrophic pastures were less damaged on stony soil but completely destroyed on deep soil, and the recovery was slow, characterised by a shift of plant species in a more eutrophic direction. Four years were not sufficient for complete recovery. Oligotrophic calcareous pastures on shallow stony soil were not damaged. Extensive breeding of pigs in mountain pastures might be harmful to plant species and vegetation, and ought to be restricted to the less sensitive plant communities, with a rotation on two to three different sites.

Kunnostusojituksen erilliskannattavuus muutamilla karuhkoilla rämeillä

Summary: Profitability of ditch-network maintenance on some oligotrophic pine mires

Reversible and irreversible pollutant-induced bacterial cellular stress effects measured by ethidium bromide uptake and efflux.

Chemical pollution is known to affect microbial community composition but it is poorly understood how toxic compounds influence physiology of single cells that may lay at the basis of loss of reproductive fitness. Here we analyze physiological disturbances of a variety of chemical pollutants at single cell level using the bacterium Pseudomonas fluorescens in an oligotrophic growth assay. As a proxy for physiological disturbance we measured changes in geometric mean ethidium bromide (EB) fluorescence intensities in subpopulations of live and dividing cells exposed or not exposed to different dosages of tetradecane, 4-chlorophenol, 2-chlorobiphenyl, naphthalene, benzene, mercury chloride, or water-dissolved oil fractions. Because ethidium bromide efflux is an energy-dependent process any disturbance in cellular energy generation is visible as an increased cytoplasmic fluorescence. Interestingly, all pollutants even at the lowest dosage of 1 nmol/mL culture produced significantly increased ethidium bromide fluorescence compared to nonexposed controls. Ethidium bromide fluorescence intensities increased upon pollutant exposure dosage up to a saturation level, and were weakly (r(2) = 0.3905) inversely correlated to the proportion of live cells at that time point in culture. Temporal increase in EB fluorescence of growing cells is indicative for toxic but reversible effects. Cells displaying high continued EB fluorescence levels experience constant and permanent damage, and no longer contribute to population growth. The procedure developed here using bacterial ethidium bromide efflux pump activity may be a useful complement to screen sublethal toxicity effects of chemicals.

Shallow-platform palaeoenvironmental conditions recorded in deep-shelf sediments: C and stable isotopes in Upper Jurassic sections of southern Germany (Oxfordian-Kimmeridgian)

A high-resolution carbon and oxygen isotope analysis of Late Oxfordian-Early Kimmeridgian deep-shelf sediments of southern Germany is combined with investigation of nannofossil assemblage composition and sedimentological interpretations in order to evaluate the impact of regional palaeoenvironmental conditions on isotopic composition of carbonates. This study suggests that carbonate mud was essentially derived from the Jura shallow platform environments and also that the isotopic signature of carbonates deposited in the Swabian Alb deep shelf indirectly expresses the palaeoenvironmental evolution of the platform. Short-term fluctuations in delta(13) C and delta(18)O are probably controlled by changes in salinity (fresh-water input versus evaporation) in platform environments. Long-term fluctuations in carbon and oxygen isotope record throughout the Late Oxfordian-Early Kimmeridgian result from the interplay of increasing temperature and decreasing humidity, which both control the trophic level. Changes from mesotrophic to oligotrophic conditions in platform environments and in the deep-shelf surface waters are inferred. During the Late Oxfordian (Bimammatum Subzone to Planula Zone), the delta(13)C curve displays a positive shift of about 1 parts per thousand, which is comparable in intensity to global perturbations of the carbon cycle. This evident isotopic shift has not been documented yet in other basinal settings. It can be reasonably explained by local palaeoenvironmental changes on the Jura platform (salinity, temperature, and nutrient availability) that controlled platform carbonate production, and the geochemistry of overlying waters. However, increasing carbonate production on the Jura platform and related positive delta(13)C shifts recorded in the Swabian Alb deep shelf are the regional signatures of climatic changes affecting other palaeogeographical domains of Europe in which the carbonate production increased throughout the Late Oxfordian. (C) 2002 Elsevier Science B.V. All rights reserved.

Surface-dependent strategies and energy flux in benthic marine communities or, why corals do not exist in the Mediterranean

Most structure-building organisms in rocky benthic communities are surface-dependent because their energy inputs depend mainly on the surface they expose to water. Two photosynthetic strategies, divided into calcareous and non calcareous algae, strict suspension-feeders and photosynthetic suspension feeders (e.g. hermatypic corals) are the four main strategies evolutively acquired by benthic organisms. Competition between those strategies occur in relation to productivity of the different species, in such a way that, for given environmental conditions, species with a higher growth (P/B ratio) would dominate. At a worldwide scale, littoral marine benthos can he considered to fit into the four fields defined by two main axes: the first, relates to productivity and relies atrophic and oligotrophic waters and the second is defined by the degree of environmental variability or seasonality (from high to low). Coral reefs (marine ecosystems dominated by photosynthetic suspension feeders) develop in the space of oligotrophic areas with low variability, while kelp beds (marine ecosystem dominated by large, non calcareous algae) are to be found only in eutrophic places with a high variability. The space of eutrophic waters with a low variability do not has specially adapted, high structured, benthic marine ecosystems, and in these conditions opportunistic algae and animals predominate. Finally, photophilic mediterranean benthos -devoid of kelps and without hermatypic corals- typifies the field of oligotrophic areas with high variability; in its more genuine aspect, Mediterranean benthos is represented by small algae with a high percentage of calcareous thallii. In all cases strict suspension-feeders compete successfully with photosynthetic organisms only in situations of low irradiances or very high inputs of POM. In its turn, Mediterranean rocky benthos, in spite of its relative uniformity, is geographically organized along the same axes. The Gulf of Lions and the insular bottoms (Balearic Islands, for example) would correspond to the extremes of eutrophic-high variability areas and oligotrophic-low variability areas, respectively. Irradiance, nutrient and POM concentration, and hydrodynamism are the three variables which mainly affect the distribution of the different surface-dependent strategies, and thus, these parameters are of paramount interest for understanding the trophic structure of Mediterranean benthic communities. In environments non limited by light, nutrient availability, defined as the product between nutrient -POM concentration and hydrodynamism, states the dominance of calcareous versus non calcareous algae. Calcareous algae dominate in oligotrophic waters while non-calcareous algae dominate in moderately eutrophic waters. In light-limited environments, passive suspension feeders (octocorallaria, gorgonians) become dominant species if POM availability is enhanced by a high hydrodynamism (strong currents); in waters with a low charge of POM organisms of other groups, mainly active suspension feeders, predominate (sponges, bryozoans, scleractiniarians). In any case, there always exists a very variable bathymetric zone, depending on light attenuation and nutrient-POM availability, where encrusting calcareous algae strongly compete with suspension feeders (coralligenous).

A comparison of the energy budgets of two freshwater Pulmonates: Lymnaea peregra (Müller) and Physa acuta (Drap.)

The energy budgets of two freshwater gastropds, Lymnae peregra and Physa acuta, were compared in similar experimental conditions (20ºC, fed ad libitum with 24h-decayed lettuce), and found to differ in several ways. 1) L. Peregra has a higher assimilation efficiency than P. acuta (72% vs 60%). 2) These species assimilate different components of the ingested food: P. acuta uses a smaller, but more energetic part (probably mainly bacteria), whereas L. peregra assimilate a larger, but less energetic part (probably mainly cellulose). 3) L. peregra allocates more of its assimilated energy to oxygene consumption and mucus production (maintenance investments), wheras P. acuta invest more in growth and reproduction (production investments). Such differences are relevant to the natural habitat of these two species: P. acuta colonizes warm, eutrophic and temporary pools, where decaying material constitue the main part of available resources, and where adult mortality is high and impredictible. By contrast, L. peregra is frequently found in colder, oligotrophic and predictible environements, where living primary producers constitute the main part of available resources, and where biotic interactions are important factors of mortality.

Recent human-induced trophic change in the large and deep perialpine Lake Lucerne (Switzerland) compared to historical geochemical variations

This study investigates the sedimentological and geochemical changes that occurred during the last 2200 years in the meromictic Lake Lucerne (Switzerland), one of the largest freshwater lakes of Central Europe. The stable isotope composition (delta C-13 and delta O-18 values) of bulk carbonates is compared to changes in grain-size distribution (clay and silt fraction), natural trace element input (titanium and thorium concentrations), and organic material abundance (C-org, nitrogen and phosphorus) and composition (C/N ratios and hydrogen and oxygen indexes). A drop in carbonate accumulation and in the delta O-18 values of sediments between ca. AD 500 and 700 followed a large and consistent rise in chemical weathering, marked by increases in the silicate-clay fraction and in crustal element concentrations. During the following millennium, there was a long-term decreasing trend in the lithogenic trace element input and in the phosphorus loading, suggesting decreasing terrigeneous input from runoff water. The major sedimentological change over the studied period occurred after ca. AD 1800 with a significant increase in the erosion-driven silt-fraction and in the sedimentation rate. During the last century, human-induced increase in nutrient input to the lake highly enhanced the accumulation of organic matter in sediment. Changes in nutrients and oxygen conditions in the hypolimnion of Lake Lucerne during the eutrophication period (i.e., the last 40 years) highly modified the geochemical fluxes compared to the relatively stable oligotrophic conditions that prevailed during the previous 2000 years. Before the 19th century, climate driven meromixis had a limited impact on the organic matter flux to the sediments, but the accumulation of carbonate considerably decreased during periods of lower mechanical erosion rates and high chemical weathering rates. (C) 2012 Elsevier B.V. All rights reserved.

Phosphorus and carbon burial during cretaceous oceanic anoxic events, links with climatic changes

According to Jenkyns (2010), oceanic anoxic events (OAE) record profound changes in the climatic and paleoceanographic state of the planet and represent major disturbances in the global carbon cycle. One of the most studied OAEs on a worldwide scale is the Cenomanian-Turonian OAE 2, which is characterized by a pronounced positive excursion in carbon-isotope records and the important accumulation of organic-rich sediments. The section at Gongzha (Tibet) and the sections at Barranca and Axaxacualco (Mexico) are located in remote parts of the Tethys, and show δ13C records, which are well correlated with those of classical Tethyan sections. Both sections, however, do not exhibit the presence of organic-rich sediments. Phosphorus Mass Accumulation Rates (PMAR) in Tibet show a pattern similar to that observed in the Tethys by Mort et al. (2007), which suggests enhanced Ρ regeneration during the OAE 2 time interval, though there is no evidence for anoxic conditions in Tibet. Ρ appears here to have been mainly driven by detrital influx and sea-level fluctuations. The sections at Barranca and Axaxacualco show that the Mexican carbonate platform persisted during this anoxic event, which allowed the evolution of platform fauna otherwise not present in Tethyan sections. The persistence of this carbonate platform close to the Caribbean Igneous Plateau, which is thought to have released bio-limiting metals, is explained by local uplift which delayed the drowning of the platform and a specific oceanic circulation that permitted the preservation of oligotrophic conditions in the area. The Coniacian-Santonian OAE (OAE3) appears to have been more dependent on local conditions than OAE2. The presence of black shales associated with OAE3 appear to have been restricted to shallow-water settings and epicontinental seas in areas located around the Atlantic Ocean. The sections at Olazagutia (Spain), and Ten Mile - Arbor Park (USA), two potential Global Boundary Stratotype Sections and Points (GSSP) sites, are devoid of organic-rich sediments and lack a δ13C positive excursion around the C-S boundary. The Gabal Ekma section (Sinai, Egypt) exhibits accumulations of organic-rich sediments, in addition to phosphorite bone beds layers, which may have been linked to an epicontinental upwelling zone and/or storm inputs. Our data suggest that OAE 3 is rarely expressed by truly anoxic conditions and seems to have been linked to local conditions rather than global paleoenvironmental change. The evidence for detrital-P being the likely cause of Ρ fluctuations during the OAEs studied here does not negate the idea that anoxia was the principal driver of these fluctuations in the western Tethys. However, an explanation is required as to why the Ρ accumulation signatures are mirrored in both oxic and anoxic sedimentary successions. 'Eustatic/climatic' and 'productivity/anoxic' models may have both operated simultaneously in different parts of the world depending on local conditions, both producing similar trends in Ρ accumulation. - Selon Jenkyns (2010), les événements anoxiques océaniques enregistrent de profonds changements dans le climat et la paléoceanographie de la planète et représente des perturbations majeures du cycle du carbone. L'un des plus étudiés à l'échelle mondiale est l'ΟΑΕ2 du Cénomanien-Turonien, qui est caractérisé par une très forte excursion positive des isotopes du carbone et une importante accumulation de sédiments riche en matière organique. La section de Gongzha (Tibet) et les sections de Barranca et Axaxcualco (Mexique) sont situées aux confins de la Téthys, et enregistrent une courbe isotopique en δ13C parfaitement corrélable avec les sections téthysiennes, mais ne montre pas d'accumulation de black shales. Le taux de phosphore en accumulation de masses (PMAR) au Tibet montre un pattern similaire observé également par Mort et al. (2007) dans la Téthys, suggérant un model de régénération du Ρ durant l'anoxie, cependant aucune conditions anoxiques régnent dans la région du Tibet. Ρ apparaît donc principalement guidé par le détritisme et les fluctuations du niveau marin. Les sections de Barranca et d'Axaxacualco montrent que la plateforme carbonatée mexicaine persiste durant cet événement anoxique, et permet le développement d'une faune de plateforme qui n'est pas présente dans les sections téthysiennes. La persistance de cette plateforme carbonatée si proche du plateau Caribéen, qui est connu pour le relâchement de métaux bio-limitant, peut être expliqué par un soulèvement tectonique local qui inhibe l'ennoiement de la plateforme et une circulation océanique spécifique qui permet la préservation de conditions oligotrophiques dans cette région. L'événement anoxique océanique du Coniacien-Santonien apparaît plus dépendant des conditions locales que pour l'ΟΑΕ2. Les black shales associés à POAE3 sont restreints aux zones situées autour de l'océan Atlantique et plus particulièrement aux eaux peu profondes et épicontinentales. Les sections d'Olazagutia (Espagne), Ten Mile Creek et Arbor Park (USA), qui sont deux potentielles sections GSSP (Sections de stratotype de limite globaux et de points), ne montre pas d'accumulation de black shales et pas de forte excursion positive en δ13C autour de la limite C-S. La section de Gabal Ekma (Sinai, Egypte) montre des accumulations de black shales, en plus des couches de phosphorites et d'accumulation d'os (« bone beds »), vraisemblablement lié à des zones active d'upwelling épicontinentale et/ou d'apport de tempêtes. Nos données suggèrent que l'OAE 3 est rarement exprimé par de vraies conditions anoxiques et semble être plus lié à des conditions plus locales que des changements paléo-environnementaux globaux, comme observés pour le Cénomanien- Turonien. Les arguments pour un modèle lié au phosphore détritique qui serait la cause des fluctuations du phosphore total durant les OAEs, n'écartent pas l'idée que l'anoxie est la principale cause de ces fluctuations dans les sections riches en matière organique de l'Ouest téthysien. Cependant une explication est nécessaire pour comprendre pourquoi la signature de l'accumulation du phosphore est semblable dans les successions sédimentaires déposées dans des conditions oxygénées et anoxiques. Les modèles « Eustatisme/Climat » et « Productivité/anoxie » ont simultanément opéré dans les différentes parties du monde dépendant de conditions locales, et ont produit des tendances similaires en accumulation de phosphore.

Reconstruction of the recent history of a large deep prealpine lake (Lake Bourget, France) using subfossil chironomids, diatoms, and organic matter analysis: towards the definition of a lake-specific reference state

This paper presents the recent history of a large prealpine lake (Lake Bourget) using chironomids, diatoms and organic matter analysis, and deals with the ability of paleolimnological approach to define an ecological reference state for the lake in the sense of the European Framework Directive. The study at low resolution of subfossil chironomids in a 4-m-long core shows the remarkable stability over the last 2.5 kyrs of the profundal community dominated by a Micropsectra-association until the beginning of the twentieth century, when oxyphilous taxa disappeared. Focusing on this key recent period, a high resolution and multiproxy study of two short cores reveals a progressive evolution of the lake's ecological state. Until AD 1880, Lake Bourget showed low organic matter content in the deep sediments (TOC less than 1%) and a well-oxygenated hypolimnion that allowed the development of a profundal oxyphilous chironomid fauna (Micropsectra-association). Diatom communities were characteristic of oligotrophic conditions. Around AD 1880, a slight increase in the TOC was the first sign of changes in lake conditions. This was followed by a first limited decline in oligotrophic diatom taxa and the disappearance of two oxyphilous chironomid taxa at the beginning of the twentieth century. The 1940s were a major turning point in recent lake history. Diatom assemblages and accumulation of well preserved planktonic organic matter in the sediment provide evidence of strong eutrophication. The absence of profundal chironomid communities reveals permanent hypolimnetic anoxia. From AD 1995 to 2006, the diatom assemblages suggest a reduction in nutrients, and a return to mesotrophic conditions, a result of improved wastewater management. However, no change in hypolimnion benthic conditions has been shown by either the organic matter or the subfossil chironomid profundal community. Our results emphasize the relevance of the paleolimnological approach for the assessment of reference conditions for modern lakes. Before AD 1900, the profundal Micropsectra-association and the Cyclotella dominated diatom community can be considered as the Lake Bourget reference community, which reflects the reference ecological state of the lake.

Polyunsaturated aldehydes from large phytoplankton of the Atlantic Ocean Surface (42ºN to 33ºS)

Polyunsaturated aldehydes (PUAs) are organic compounds mainly produced by diatoms, after cell wounding. These compounds are increasingly reported as teratogenic for species of grazers and deleterious for phytoplanktonic species, but there is still scarce information regarding concentration ranges and the composition of PUAs in the open ocean. In this study, we analyzed the spatial distribution and the type of aldehydes produced by the large-sized (>10 μm) phytoplankton in the Atlantic Ocean surface. Analyses were conducted on PUAs released after mechanical disruption of the phytoplankton cells, referred to here as potential PUAs (pPUAs). Results show the ubiquitous presence of pPUA in the open ocean, including upwelling areas, as well as oligotrophic gyres. Total pPUA concentrations ranged from zero to 4.18 pmol from cells in 1 L. Identified PUAs were heptadienal, octadienal and decadienal, with heptadienal being the most common (79% of total stations). PUA amount and composition across the Atlantic Ocean was mainly related to the nitrogen:phosphorus ratio, suggesting nutrient-driven mechanisms of PUA production. Extending the range of trophic conditions considered by adding data reported for productive coastal waters, we found a pattern of PUA variation in relation to trophic status.

Community replacement of neritic carbonate organisms during the late Valanginian platform demise: A new record from the Provence Platform

The Valanginian is marked by a major platform demise inducing a hiatus in the northern Tethyan neritic carbonate record from the top of the lower Valanginian to the lower Hauterivian. New biostratigraphic and chemostratigraphic data from the Ollioules section (Provence Platform, southern France) are presented here, demonstrating that a large part of the upper Valanginian is preserved in an inner platform environment. The thick, upper Valanginian, aggrading carbonate succession is observed in an aborted rift domain, implying relatively low subsidence. In this context, a relatively long-term sea-level rise was required to sustain a keep-up style of carbonate production. Like the Apulian Platform, the remarkable preservation of the Provence Platform may have been favored by its remoteness from terrigenous source areas, as suggested by the low clastic inputs and low P-accumulation rates. Two main biotic community replacements are observed in Ollioules. The first saw the development of abundant microbialites and algae at the onset of the late Valanginian. A Tubiphytes concentration occurred during the coolest climatic conditions and the transition towards arid conditions, whereas the subsequent Lithocodium-Bacinella and orbitolinids assemblages developed under low nutrient conditions during a warmer interval. Both assemblages may have been triggered by increased alkalinity. The second community replacement saw the installation of coral- and rudist-dominated communities during the latest Valanginian to early Hauterivian. They indicate a change to oligotrophic, open marine conditions. Six medium-scale sequences have been defined in Ollioules, indicating short-term transgressive-regressive trends superimposed on a long-term transgression. Low nutrient inputs and relatively low subsidence in an aggradational context may explain the survival of the isolated Provence Carbonate Platform during a time of widespread drowning episodes and platform demise in the northern Tethyan domain. (c) 2012 Elsevier B.V. All rights reserved.

Benthos of Spanish lakes and reservoirs

A summary of the results from the study of benthos of lakes and reservoirs in Spain is provided, with a list of the species found to date. Spanish natural lakes are smaller than European lakes; the largest is Lake Sanabria, of glacial origin, which is 3 Km long and half a kilorneter wide. Many are very small and situated in the mountains; more than 200 hundred have been recorded in Spain, but only in Lake Sanabria and Lake Banyoles have the benthos been studied. Lake Sanabria is a cold oligotrophic, monomictic lake with oxygen always present in the deepest zones. Its fauna is similar to that of other central European lakes, with Mici.opsectra c.orztractu (a chironomid) as the dominan1 species. Lake Banyoles is a karstic, monomictic and multibasin lake; despite the low primary productivity, due to the abundante of sulphate in the water, the allochthonous inflow of organic matter and the inflow of water from bottom springs, the profunda1 environinent is very stressing for benthic fauna. Very low oxygen concentrations and high sulphide content in the water and sediments dueto meromixis mean that only the larva of the dipteran Chaohoi.lcs flai7icans was present in one of the 5 basins of the lake. In other basins, when oxygen is available (no meromixis), the fauna is similar to that of tlie inineralized lakes of the Aegean region and some lakes in central Italia. On the other hand, preliminary data from the Pyrenean lakes and from Sierra Nevada ponds reveal no differences with northern cold lakes. Tlie largest lakes in Spain are the reservoirs. There are nearly 1000 and data are available on 100 of them, including the kargest. In addition to oxygen and sulphide content in the bottom waters, water level fluctuation and high sedimentation rates are disturbance factors that prevented the organization of the community. Allochthonous inputs of organic matter are also an important factor both in the reservoirs and also in the small, oligotrophic lakes like Banyoles and Sanabria. As a result the meiofaunal loop is very important in many of the Spanish water bodies . For this reason the natural lakes and reservoirs of Spain are dominated by Oligochaeta, small crustaceans and the microcarnivore chironomids (such as Procladi~ls, Cladopelma and Mi(,rnc.hil-onnmus) that feed on these meoifaunal elements. The phytophagous chironomids, like Chironomus, are only abundan1 in the shallow areas of mesotrophic and eutrophic reservoirs. This situation makes it difficult to apply the typological system of SAETHER which predicts with some confidence only the benthic communities of Spanish natural lakes above 1500 m in the Pyrenees or the ponds above 2000 m in Sierra Nevada mountains. Higher temperatures (which originate a longer stratification period), the presence of sulphate in the waters of the eastern part of Spain and high inputs of sediments and allochthonous organic matter seem to be the factors that originated the differences between the benthic profunda1 faunas of Spanish lakes and reservoirs and those of the temperate lakes of north and central Europe.