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In an applied research project on the development of a pulsed microwave sulfur lamp prototype of 1 kW, we have discovered an amazing phenomenon in which the plasma forms a ball staying at the center of the bulb despite gravity, thus protecting the glass from melting. In this paper, it is shown that this results from an acoustic resonance in a spherical mode. Measurements of the plasma response to short pulses are presented showing beats at the spherical resonance. It is demonstrated that the beats could result from the simultaneous excitation of two normal modes with a frequency difference of approximately 1%. One of the two frequencies matches precisely the microwave pulses repetition, a little below 30 kHz. Thus this one is due to a forced oscillation, whereas the other one is due to a free oscillation. The phase velocity of sound was calculated as a function of temperature in order to find the series of temperatures at which a resonance would occur if the bulb were an isothermal solid sphere. The mean temperature inside the actual bulb was determined from the only doublet of this series, that has characteristic frequencies close enough to cause the observed beats. In addition, one of these two modes has a spherical symmetry that can explain the plasma ball formation. The obtained mean temperature is consistent with the direct measurements on the bulb surface as well as with the temperature in the core of a similar plasma found in the literature. We have also proposed a model of the resonance onset based on the acoustic dispersion and the sound amplification due to electromagnetic coupling.

International audience; For several years, major exoskeletal anomalies in two species of European estuarine shrimp of the genus Palaemon have been observed. The phenomenon has been described for P. longirostris and P. macrodactylus in the Gironde estuary (France) and affects all stages of the species’ life cycles: from juvenile to adult and in both wild and laboratory-reared individuals. In the present follow-up study, the effects of these anomalies in larval stages of the two species were investigated. Regular sampling across most of the saline estuary confirmed that the post-larval stages of both species were affected. The morphological anomalies (MA) observed in the larvae were of the same type as those described for the adult stages, although more limited and primarily affecting the rostrum and cephalothorax. Larvae were reared from hatching to the post-larval stage, and the appearance of the aforementioned anomalies was observed as early as in Zoea I. This was the case even in larvae whose mothers did not exhibit anomalies. For both species, the most advanced stage was the one most affected. Also, in the rearing experiment, a greater proportion of the P. macrodactylus larvae was affected than those of P. longirostris. No significant effects ofMA on size or growth rate were demonstrated for larvae of either species in this study. Nevertheless, it provides data on larval growth that were not previously available. Further rearing experiments should be considered in order to explore any demonstrable effects of these morphological anomalies.; Deux espèces de crevettes estuariennes européennes du genre Palaemon sont affectées depuis plusieurs années par des déformations importantes de l’exosquelette. Ce phénomène a été décrit pour les espèces P. longirostris et P. macrodactylus de l’estuaire de la Gironde (France) et porte sur les stades juvéniles à adultes, provenant du milieu naturel et également suivis en élevage. Dans cette étude complémentaire, les stades larvaires de ces deux espèces sont étudiés. L’échantillonnage régulier des larves sur la majeure partie de l’estuaire salé a confirmé une atteinte des stades postlarvaires des deux espèces et ceci dans des proportions significatives. Les déformations observées sur les larves sont du même type que celles décrites pour les stades adultes, avec toutefois une expression morphologique plus restreinte touchant principalement le rostre et le céphalothorax. L’élevage des larves, depuis l’éclosion jusqu’au stade post-larve, a permis d’observer l’apparition de ces déformations dès le stade Zoé I, même chez les mères pourtant dépourvues de déformation. Chez les deux espèces, les stades les plus avancés sont les plus affectés. Aussi, nous avons montré, lors de l’élevage au laboratoire, qu’une plus grande proportion de larves de P. macrodactylus était affectée par rapport à P. longirostris. Aucun effet significatif des anomalies morphologiques sur la taille ou le taux de croissance des larves n’a pu être démontré pour les deux espèces. Cependant, des données sur la croissance larvaire des deux espèces sont apportées. D’autres expériences d’élevage en laboratoire devraient être envisagées afin d’explorer les effets de ces anomalies morphologiques.

Aqueous extraction of sunflower seeds and rapeseeds with mixtures of hydrolytic enzymes were carried out in 2-L reactor and uronic acid and reducing sugar concentrations were measured during digestion. The goal of the study was to determine if a correlation between those concentrations and free oil exists that would allow predicting free oil release from aqueous solution measurements. Similar concentrations of 4.5 and 4.7 g/L of reducing sugars and 43 and 55 g/L of uronic acid were found for sunflower and rapeseeds, respectively. The corresponding yields of free oil from the two seeds were 83% and 16% of total oil, and this difference was due to the formation of an emulsion in the rapeseed dispersion. Therefore, measurement of reducing sugars or uronic acid concentrations are not sufficient to predict a release of free oil from all oil seeds.

Palaemon macrodactylus, an exotic shrimp native to the northwest Pacific Ocean, has been recorded in many estuaries along the Atlantic coast of Europe since the late 1990’s. In this study, a regular monthly survey, held since 1992 of the middle section of the Gironde estuary, revealed this species’ rapid and full colonization of the system since its appearance in samples during summer 1998. In the Gironde, the population of P. macrodactylus is self-sustaining and the species is now established there. Our study highlights two important elements that might explain its successful colonization: its ability to quickly invade a niche under-exploited by the similar native species and its greater reproductive output. The species’spatio-temporal distribution in relation to environmental variables was studied and compared with that of the native species P. longirostris. The exotic species was shown to mainly inhabit the polyhaline and mesohaline section of the estuary, which are sections generally under-exploited by females of the native species. Furthermore, fecundity analysis and larval abundance survey results revealed a higher potential fecundity and a greater proportion of larvae and juvenile stages for P. macrodactylus in the natural environment.

Our study details key parameters of the reproductive biology of the shrimp Palaemon longirostris, e.g., spatio-temporal variation of the sex-ratio, migration pattern, breeding period, fecundity, size at the onset of maturity, and egg volume. While P. longirostris population was very little studied in its distribution area, our fist aim was to complete that lack of knowledge and to provide a basis for understanding its population dynamic. Finally, our results were also compared with those reported from different studies on this species within Europe, in order to test the reported latitudinal cline of reproductive parameters in decapods. During a one-year survey, P. longirostris have been collected monthly across a large part of the Gironde estuary (France). Biometric measurements and determination of sex and maturation stages were carried out on preserved subsamples. Eggs were counted and measured for ovigerous females. In the Gironde, the breeding period was restricted from March to July, and rel...

Enzymic synthesis of fructose esters was studied under reduced pressure. Different acyl donors were tested, and immobilized Candida antarctica lipase was used as biocatalyst. Influences of pressure, nature of the acyl donor, molar ratio sugar/acyl donor were investigated. Pressure had the greatest influence. At 200 mbar, more than 90% of fructose was acylated compared to 50% under atmospheric pressure. This is explained by the evaporation of reaction by-product (methanol or water) that shifted the equilibrium. C. antarctica lipase catalyzed sugar ester synthesis very efficiently using rapeseed oil as acyl donor. Moreover, synthesis performed with an equimolar mixture of both substrates gave promising results. Although the reaction rate was slower than synthesis performed with an excess of fatty acid, fructose monooleate concentration was still high (44 g l−1 instead of 56 g l−1) and the residual acyl donor concentration was very low. Downstream processes for the recovery of pure fructose monooleate were simplified in this case.

Rougier, T., Lambert, P., Drouineau, H., Girardin, M., Castelnaud, G., Carry, L., Aprahamian, M., Rivot, E., and Rochard, E. 2012. Collapse of allis shad, Alosa alosa, in the Gironde system (southwest France): environmental change, fishing mortality, or Allee effect? – ICES Journal of Marine Science, 69: 1802–1811. At the end of the 20th century the allis shad population in the Gironde was the largest in Europe. During the first decade of the 21st century, catches declined dramatically by two orders of magnitude, and a fishery moratorium was implemented in 2008. This deterioration in the status of the stock was confirmed by three independent assessments (abundance of juveniles and of potential and effective spawners). Three hypotheses on the cause of the collapse were examined: (i) an environmental change in freshwater and/or in the estuary; (ii) an increase in marine and/or estuarine mortality; and (iii) the presence of an Allee effect. Changes in flow, temperature, and water quality over the period were inconclusive, but remain a possible causative factor. The instantaneous rate of marine (mean: 0.7 year−1, s.e. 0.1 year−1) and estuarine (mean: 2.6 year−1, s.e. 0.1 year−1) mortalities showed no trend between 1991 and 2003. Nevertheless, the past high estuarine (fishing) mortalities combined with a demographic Allee effect in the reproduction dynamics could explain the population collapse and hamper the stock recovery under the moratorium. This conclusion is, however, tentative as it was not possible to prove the presence of the density-dependent mechanism associated with the demographic Allee effect.

International audience; In view of the European Union (EU) requirements to diagnose and monitor the ecological status of estuarine ecosystems using fish-based indicators, we assessed the effect of the sampling protocol in meeting these objectives. Using three different sampling surveys, we evaluated the impact of four components, i.e. sampling gear, spatial extent, length of time series and temporal window analysed, on the detection of ecological trends in the Gironde estuary, estimated by different types of fish-based indicator. For each component, temporal trends of several fish-based indicators were analysed and for the final diagnosis these temporal trends were combined. The results showed that sampling gear had significant effects on the assessment of ecological trends, as it was determinant in the functional description of the fish community and also in the final diagnosis obtained. When on the one hand the lower estuary and the estuarine plume and on the other hand the upper-middle estuary, were considered, we were able to visualise the spatialized functional organization of the estuary and to assess its ecological status more precisely. Increased length of marine fish species in the lower estuary and increased delta diversity in the middle estuary suggested that larger individuals of marine species had moved from outside the estuary to the middle estuary. These trends, associated with the spatial change in environmental conditions (salinity and temperature) in the Gironde estuarine ecosystem may be indicative of a functional shift, from diadromous to marine migrant-dominated assemblages in that ecosystem. We hypothesized that the Gironde estuary was actually more suited to providing nursery habitats for the juveniles of marine migrants and marine stragglers than providing migratory corridors and nursery habitats for species seeking low salinities for a period in their life cycle. The present study supported the hypothesis of a marinisation of the Gironde estuary. When the four components of the protocol, i.e. gear, space, and time (length and temporal windows of time series), were compared this study also suggested that fish length and notably the “temporal window” on which the analyses were based were the most “sensitive” components of the sampling protocol when assessing fish ecological trends in estuaries. It clearly underlined the fact that long time series were essential for evaluating changes in the ecological functioning of these estuarine ecosystems.

International audience; Spanning 20 years (1979–2007), this study is the longest time series pertaining to the resident shrimp species Palaemon longirostris in a European estuary. Data from monthly faunal surveys undertaken across the middle part of the Gironde estuary from April 1979 along with data from a statistical analysis of the commercial catches throughout the entire estuary and river were considered in order to explore their inter and intra-annual variability and long-term trends. Long-term densities series as well as environmental series (salinity, discharge, temperature and NAO) were decomposed and the effects of environmental variables were also examined using statistical models (GAMs). This revealed important spatio-temporal variability and a significant overall decrease in abundance of this species in the Gironde estuary since the beginning of the 1980s. This longterm decrease in abundance corresponded significantly to long-term decreases in both discharge and the NAO index, as well a long-term increase in temperature and salinity in the middle part of the estuary. However, models showed that environmental factors explained only a small part of the variability. The upstream shift of the population highlighted in this study, probably due to the intrusion of marine waters into the middle section of the estuary, may also have contributed to its decrease in abundance. Inter-annual variability of densities was also significantly linked with inter-annual fecundity fluctuations, and a significant decrease in both mean female size and fecundity was shown for preserved samples from 1992. Moreover, the breeding period has been temporally stretched out and began earlier in more recent years, potentially due to the increase in spring temperature.

International audience; Chitosan particles were functionalized with ferulic acid (FA) and ethyl ferulate (EF) as substrates using laccase from Myceliophtora thermophyla as biocatalyst. The reactions were performed with chitosan particles under an eco-friendly procedure, in a heterogeneous system at 30 degrees C, in phosphate buffer (50 mM, pH 7.5).The FA-chitosan derivative presented an intense yellow-orange color stable while the EF-chitosan derivative was colorless. The spectroscopic analyses indicated that the reaction products bound covalently to the free amino groups of chitosan exhibiting a novel absorbance band in the UV/Vis spectra between 300 and 350 nm, at C-2 region by the duplication of C-2 signal in the C-13 NMR spectrum, via Schiff base bond (N=C) exhibiting novel bands in the FT-IR spectrum at 1640 and 1620 cm(-1). Additionally, antioxidant capacities of chitosan derivatives showed that the chitosan derivatives presented improved antioxidant properties, especially for FA-chitosan derivative (EC50 were 0.52 +/- 0.04, 0.20 +/- 0.02 mg/ml for DPPH center dot and ABTS(center dot+) scavenging, respectively). (C) 2011 Elsevier Ltd. All rights reserved.

Growth and morphometric characteristics of the caridean Palaemon longirostris Milne Edwards, 1837, were studied in the Gironde estuary population. Their first comparison with earlier studies in this estuary and in the Mira River provided the first step towards developing environmental indicators using a species represented at the European scale. Mean individual growth was modelled for each sex using the classical von Bertalanffy growth model and the seasonal model that integrates periods of lower growth. Morphometric relationships, in particular the spatio-temporal variability in the relative weight of individuals, were explored according to sex and physical parameters of the environment. Females grew to larger size than males. Growth estimates showed the importance of accounting for the seasonal variations to evaluate non-biased growth. Sex and the presence of eggs for females showed significant interaction within the carapace length - total length relation as well as for the carapace length - weight relation. Body condition was significantly greater in spring and summer for both sexes and was significantly correlated with salinity, but only for males. Comparison with the Mira River was limited to the range of data available. However, the population of the Gironde estuary, being similar in both recent and historical studies, showed morphometric differences when compared to that of the Mira River. Indeed, the growth performance index was higher in the Gironde estuary and likely linked to temperature constraints. Conversely, differences between males and females were much higher in the Mira River than in the Gironde estuary.

A highly sensitive, rapid LC–APCI–MS method for identification and quantification of mono and disaccharides in simple or complex aqueous phase has been developed. This original method is easy to use, no derivation and no post-column injection are needed. The separation is performed with a hydrophilic amino interaction (HILIC) column allowing high-throughput analysis with analysis times of 15 min for monosaccharides to 22 min for disaccharides. The development of the method carried out with 9 standard saccharides allowed to point out a dynamic range from 0.1–25.6 to 1–256 μg mL−1 depending on the considered sugar. Next, the method was validated on saccharides at known concentrations in water and on 2 real samples: orange juice and aqueous phase obtained after enzymatic hydrolysis of sunflower seeds.

The 4-chloro5-nitropyrimidines 2 and 5 upon treatment with sodium azide, rearrange to give nitromethylene tetrazole 4 and 6 respectively.

Shrimps of the Palaemon genus have been affected for over 15 years by a problem of exoskeletal deformities, particularly in the Gironde estuary (France). Given the large numbers of individuals affected, this study focus on a better description of the phenomenon to estimate its impact on shrimps. This study, on Palaemon longirostris and Palaemon macrodactylus, is based on samples collected from 1992 to 2007 in the Gironde estuary with particular focus on the year 2007, and on a 4-month rearing of adult shrimps in experimental system. The different approaches reveal the relative persistence of the phenomenon since 1992 (with on average more than 58% of ovigerous females affected) and its persistence during shrimp life cycle, even after several moults. Important consequences associated with deformities at the individual level were demonstrated: a higher mortality, a lighter egg mass and a lower weight for a given size. The presence of exoskeletal deformities is now a major phenomenon, with profound effects on individual shrimps, and which may in turn have an important impact on the survival of the P. longirostris population in particular.

Despite the high complexity and variability of estuaries, these ecosystems are very productive and play an important role in fish feeding. This paper constitutes a preliminary investigation to test how fish optimize the use of the available trophic resources, by studying trophic preference variability and feeding strategies of some pelagic and demersal fish in the Gironde estuary (southwest France). Fish and their prey were collected approximately every two months from July 2003 to June 2004 in the upstream area of the saline estuary. Stomach content analyses were realized to describe the variability of fish feeding according to their size and the time of year. Intra- and interspecific food niche overlap was evaluated using Schoener's index and a cross-calculation method was used to highlight the general fish trends in predation strategy. Stomach content results showed interspecific and intraspecific variability in fish feeding, which can be explained by their different or evolutionary ecomorphology. Their diets are composed mainly of zooplankton and hyperbenthic crustaceans with temporal variations in the consumed taxa. Optimization of the available trophic resource use, a key element in estuarine resilience, is thus possible due to the temporal adaptation of this structural trophic web. However, in spite of their temporal adaptation capacity, most fish species exhibited a specialist feeding strategy. This result was not expected, especially in the high turbidity of the Gironde estuary; the loss of one of these species could affect the fish trophic web structure and hence the resilience of the system.

The invasive oriental shrimp Palaemon macrodactylus Rathbun, 1902 has considerably extended its distribution in transitional waters along the Atlantic and Channel coasts of France during the period 2007-2010. The most probable method of a primary introduction of this species is ballast waters, but passive transport by water currents is also a possible mechanism of colonization (secondary introductions). Palaemon macrodactylus is a powerful invader of transitional waters and these new populations should be monitored in the future to assess any consequences to native species.

The Water Framework Directive requires the assessment of the ecological status of transitional waters considering the fish component. An original methodology, based on a pressure-impact approach, was established to develop a multimetric fish-based index to characterize the ecological quality of French estuaries. An index of contamination, based on the chemical pollution affecting aquatic systems, was used as a proxy of anthropogenic pressure. The fish metric selection was based on their response to disturbances tested via statistical models (generalized linear models) taking into account sampling strategy and estuarine features. Four metrics, for which discriminating responses to level of pressure were demonstrated, were retained to constitute the estuarine multimetric fish index. This new tool appeared particularly relevant to detect the contamination effects on fish communities in estuaries. It could help managers to take decisions in order to maintain or reach the good status required by the Water Framework Directive for 2015.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE; National audience; Présentation des principaux résultats et préconisations pour l'action publique. Mise en application de l'indicateur et évolution de cet indicateur poisson depuis l'achèvement du projet Liteau II.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE; International audience; Estuaries serve as nursery grounds for many marine fish species. However increasing human activities within estuaries and surrounding areas lead to significant habitat loss for the juveniles and decrease the quality of the remaining habitats. This study is based on the data of 470 beam trawls from surveys that were conducted in 13 French estuaries for the purpose of the European Water Framework Directive. It aimed at testing the effects of anthropogenic disturbances on the nursery function of estuaries. With a multispecific approach based on ecological guilds, two fish metrics, abundance and species richness of Marine Juvenile migrant fishes, were used as proxies for the estuarine nursery function. Indices of heavy metal and organic contaminations were used to estimate anthropogenic disturbances impacting these estuaries. Fish metrics were described with statistical models that took into account: (a) sampling protocol, (b) estuarine features and (c) contamination. The results of these models showed that the fish metrics highly depend on the sampling protocol, and especially type of gear, depth and salinity, which highlights the necessity of considering such metrics at the sampling (trawl haul) scale. Densities and species richness of Marine Juvenile fishes appeared to be strongly and negatively correlated to contamination indices. These results are consistent with the hypothesis that human disturbances impact the nursery function of estuaries. Finally, the densities of Marine Juvenile migrant species appeared as a potential robust and useful fish indicator for the assessment of the ecological status of estuaries within the Water Framework Directive.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE; International audience; This study has analysed for the first time fish composition and assemblage structures of three small macrotidal estuaries of the Eastern English Channel (EEC) and has explored the influences of 19 biotic and abiotic variables on the fish assemblages. Fish from Canche, Authie and Somme estuaries were collected during spring (June 2006 and May 2007) and autumn (September 2006) along the estuarine gradients using a 1.5 m beam trawl. Using identical sampling protocols, the study also analysed and compared for the first time taxonomic and functional aspects of the fish assemblages in 15 estuaries located along the Atlantic and English Channel coasts. SIMPER analysis showed high similarities in fish assemblages in the three EEC estuaries and during either spring or autumn periods. However, intraestuary similarities were relatively low, indicating that fish assemblage structures (species richnesses or abundances) were more variable within the estuary (salinity gradient) than between estuaries and/or seasons (spring vs autumn). Although numerous environmental variables were included in the study, only 47% of the variability observed in the fish distribution was explained. Fish spatial variations in the EEC estuaries are mostly driven by abiotic variables as opposed to biological interactions. As indicated by CCA, salinity and muddy sediments were the two most important factors structuring the fish assemblages. The macrobenthos being very abundant in the EEC estuaries (5801121 ind. m2), the availability of potential prey is probably not a limiting factor in the utilization of estuaries by fish. Contrary to the majority of French estuaries dominated by estuarine species (ES), the fish assemblages of the EEC estuaries are clearly dominated by marine migrant (MM) species (65% on average) with high abundance of juveniles (mostly young-of-the-year). Cluster and SIMPROF's analyses distinguished the functional structure of the 15 estuarine fish assemblages into different clusters. The three EEC estuaries form a similar group with the largest French estuaries (Seine, Loire and Gironde). The latters were characterized by higher MM/ES ratio compared to the other French estuaries. Freshwater flow and tidal range were the main factors associated with this group of estuaries. The results of the present study do not indicate that anthropogenic impacts (chemical contamination) on estuaries affect their ecological functioning as described by the fish ecological guilds. It is suggested that the guild approach may not be useful to provide valuable information on the ecological status of estuaries.

International audience; For the first time, the presence of a ferulic acid esterase (FAE) was demonstrated in Streptomyces ambofaciens. This extracellular enzyme was produced on a range of lignocellulosic substrates. The maximal level of activity was detected in the presence of either destarched wheat bran or oat spelt xylan as the sole carbon source. We found that 1% (m/ v) of destarched wheat bran was the optimal concentration to induce its production. With this inducer, no ferulic acid dimers were released from the cell wall by the produced FAE. Interestingly, rape cattle cake (Brassica napus), which does not contain esterified ferulic acid, was also shown to induce the production of the FAE from S. ambofaciens. The FAE was partially purified from the culture supernatant. The purified enzyme was optimally active at pH 7 and 40 8C. The substrate specificity of the FAE from S. ambofaciens was investigated: the highest activity was determined with methyl p-coumarate, methyl ferulate, and methyl cinnamate. Furthermore, the FAE required a certain distance between the benzene ring and the ester bond to be active. According to these biochemical characteristics, the FAE from S. ambofaciens has been classified as a type B FAE.

National audience; L'indicateur poisson pour les eaux de transition a été développé en recherchant les réponses des peuplements de poissons aux différents types de pression qui s'exercent sur ces masses d'eau. Un groupe d'expert a établie les réponses attendues pour une sélection de métriques candidates face à une augmentation des pressions. A partir de cette liste de 32 métriques candidates, des tests de corrélation ont été réalisé pour éliminer les redondances. Nous avons ensuite testé à l'aide de modèles linéaires généralisés les réponses de chaque métrique restante en fonction des niveaux de pression pour retenir à la fin 4 métriques de densité pour les estuaires qui répondent à des pressions de pollution et 3 métriques pour les lagunes qui répondent à des pressions d'occupation des sols rivulaires. Les métriques actuellement retenues sont robustes et des seuils de classe peuvent être calculés. Cependant, la référence de très bon état reste toujours à modéliser. Le faible nombre de métriques nous semble apporter un risque supplémentaire de mauvais diagnostic et des métrique complémentaires sont actuellement recherchées pour réduire les risques de classement erroné.

National audience; Les masses d'eau de transition (MET estuaires et lagunes), telles que définies par la Directive Cadre sur l'Eau (DCE directive 2000/60/CE), assurent une fonction de nourricerie pour de nombreuses espèces de poissons marins. Elles sont néanmoins soumises à de fortes pressions anthropiques qui conduisent à la destruction de l'habitat et à la dégradation de sa qualité, et mettent en danger le renouvellement des espèces concernées. Pour le suivi de l'état des MET, la DCE institutionnalise, entre autres, l'utilisation d'indicateurs ichtyologiques. Un projet du programme national Liteau 2 du Ministère en charge de l'Environnement vise notamment à développer des indicateurs « poissons » pour ce suivi. Dans ce cadre un inventaire ichtyologique standardisé est en cours de réalisation sur une large gamme de MET en France. A partir des données ichtyologiques issues de cet inventaire, plusieurs métriques représentatives de la fonction de nourricerie des MET ont été définies ; elles sont basées sur une approche multispécifique via l'utilisation de guildes écologiques. Ensuite, la sensibilité de ces métriques au protocole de récolte des données ainsi qu'à des facteurs environnementaux a été évaluée. D'autre part, des indicateurs de contamination chimique des MET ont été développés. Enfin, des modèles pressions / impacts ont été réalisés pour déterminer l'effet des pressions anthropiques sur la fonction de nourricerie des MET. Ce travail montre qu'avec un protocole standardisé d'inventaires ichtyologiques, une bonne couverture spatiale et la prise en compte de facteurs environnementaux, il est possible de mettre en évidence les effets négatifs conséquents de certains contaminants sur la fonction de nourricerie des estuaires. Ces contaminants provoquent la diminution de l'abondance et du nombre d'espèces de juvéniles qui utilisent les estuaires comme nourricerie. Finalement, une méthodologie pour définir des normes et des points de référence a été développée afin de créer des indicateurs ichtyologiques liés à la fonction de nourricerie des MET qui soient opérationnels dans le cadre de la DCE.

International audience; Carbon and Nitrogen stable isotopes and stomach contents analyses were used to investigate an estuarine fish food web and identify the contribution of these two methods to the knowledge and understanding of the food web's structure and its functioning. The nine most abundant fish species during the warm period in the Gironde estuary (southwest France, Europe) are examined. Observation of the stomach contents reflects a variety of feeding modes between fish species that consume a diverse assortment of prey, with limited dietary overlap. Nevertheless, when regarding the whole fish community, few prey species dominate the stomach contents. Nitrogen isotope ratios indicate a high intraspecific variability inducing an interspecific covering of the signatures. However, a tendency to d15N enrichment according to the trophic position of the species studied was observed. Fish assemblages show a trend towards enrichment of their carbon isotopic signatures from the upper estuary (20.8 1.8&) towards the lower estuary (18.3 1.6&). But whatever the capture zone considered, most of the individual d13C values for each fish analysed are comprised between 22 and 16&. Only few specimens, belonging to migratory amphihaline species, have significantly lighter values. The stomach contents method of analysis has the advantage of giving an initial view of the ichthyological trophic structure of the system by describing the food relations between a fish species and its prey. From these results, hypotheses can be drawn about the network's functioning, suggesting a sharing of resources between species and a wasp-waist' control of this estuarine food web. The stable isotope analysis method enables us to improve our structural knowledge by positioning the different species in a food web, with their position being determined by the number of energy transfers (analysis of d15N). Conversely, in environments as complex and changing as estuaries, it appears difficult to precisely identify and quantify the sources of the organic matter at the base of the fish estuarine food webs using analysis of d13C isotopic signatures. Nevertheless, the results obtained by using these two methods in parallel suggest that more detailed functional ecological studies could be carried out in future.

International audience; This study aims at analyzing morphological deformities in Palaemon population in the Gironde estuary. The most frequent abnormalities affected cephalothorax and rostrum, and to a lesser extent scaphocerites and uropods (with spectacular dissymmetry), whilst only few cases of antenna and telson deformities were observed. References on morphological abnormalities of crustaceans can be found in the literature, but dealing with only isolated individuals and not for Palaemonids species, whilst in the Gironde estuary up to 40% of adult individuals are affected. Deformities likely depend on water pollutants, among others heavy metals and PCBs. This hypothesis should be tested considering other European estuaries with similar features.

International audience; During summer 2006, the first specimens of the oriental shrimp Palaemon macrodactylus Rathbun, 1902 were identified in the Gironde Estuary (France). These specimens, which included ovigerous females, are confirmed present throughout a large part of the estuary, in large numbers compared to the local resident shrimp Palaemon longirostris Milne Edwards, 1837. Native to Asia, P. macrodactylus was first introduced in California (USA) in the 1950s, before being detected recently within several European estuaries, e.g. the Guadalquivir Estuary (Spain) and the Orwell Estuary (England). According to these records, P. macrodactylus appears to be a strong invader, able to colonise a wide geographical range.

International audience; The Gironde estuary, one of the largest in Europe, is considered as a reference ecological system, with all the western European diadromous fish species present. The national biomonitoring program on the coastal marine environment has revealed since 1979 severe metal pollution (mostly cadmium [Cd]) in oysters collected from the estuary. No data are available on metal contamination levels in fish, despite their ecological and economic importance. We present the results from a detailed study based on 4 metals (Cd, zinc [Zn], opper [Cu], and mercury [Hg]) measured in 4 organs (gills, dorsal skeletal muscle, liver, and kidneys) from 8 fish species illustrating several ecological combinations: European eel (Anguilla anguilla), twaite shad (Alosa fallax), bass (Dicentrarchus labrax), meagre (Argyrosomus regius), flounder (Platichthys flesus), thinlipped grey mullet (Liza ramada), sole (Solea vulgaris), and Canary drum (Umbrina canariensis). The results show very marked differences between species and organs, as well as very significant variations between the 4 metals. Although metal concentrations measured in fish muscle are low, except in the case of Hg for the A. fallax, high levels of Cu and Cd were observed in the kidneys and livers of L. ramada and A. anguilla. A multifactorial analysis based on rank ordered metal concentrations for the 8 fish species clearly shows 4 clusters of species assigned to the different degrees of metal contamination, from the lowest contaminated (A. regius, D. labrax, S. vulgaris, and U. canariensis) to the most contaminated group (L. ramada). The most contaminated species (L. ramada, A. anguilla, and P. flesus) are characterized by long residence times in the estuary, between 3.5 and 14 yr. For L. ramada, biofilms with high metal storage capacities would be the principal uptake route; the two other species are benthic with a carnivorous regime. Comparisons between our data and four estuaries (Seine, France; Mersey, U.K.; La Plata, Argentina; Guadalquivir, Spain), on a limited number of common species, metals and fish organs, clearly reveal higher Cd bioaccumulation levels in the Gironde estuary.

The 4-chloro5-nitropyrimidines 2 and 5 upon treatment with sodium azide, rearrange to give nitromethylene tetrazole 4 and 6 respectively.

L'ichtyofaune estuarienne est à la fois diverse et hétérogène. Ses grands traits écologiques rendent difficile la mise en place d'un programme de suivi. Un échantillonnage rétrospectif des données de chalutage acquises par le Cemagref depuis 1994 sur l'estuaire de la Gironde nous a permis de définir les bases d'un protocole de suivi de la richesse ichtyofaunistique en milieu estuarien compatible avec les objectifs de la Directive cadre européenne sur l'eau (DCE) adoptée en juin 2000. Un programme d'échantillonnage situé entre vingt et trente traits de chalut à deux périodes de l'année, en automne et au printemps, apparaît suffisant pour avoir une bonne représentativité de la diversité spécifique sur l'année. Nous préconisons d'adopter une stratégie qui consisterait à effectuer trois années d'échantillonnage consécutives tous les trois, quatre ou cinq ans. / The estuarine fish fauna is both diverse and heterogeneous. Its main ecological features make difficult the development of a monitoring program. A retrospective sampling of the trawling data collected by the Cemagref since 1994 allowed us to define the basis for a monitoring protocol of the estuarine fish species diversity consistent with the objectives of the new Water Framework Directive (WFD). A sampling effort between 20 and 30 trawl hauls twice a year, in autumn and spring, can be sufficient to correctly describe the species richness of the year. We recommend to adopt a strategy that would consist to sample 3 consecutives years every 3, 4 or 5 years.

La Directive Cadre sur l'Eau (DCE directive 2000/60/CE), instaure l'utilisation d'indicateurs ichtyologiques pour le suivi de l'état écologique des masses d'eau de transition (MET estuaires et lagunes). A partir de données ichtyologiques issues de campagnes DCE réalisées sur 13 estuaires en France métropolitaine, un premier travail, basé sur une approche multispécifique via l'utilisation de guildes écotrophiques, a montré que certaines métriques liées à la fonction de nourricerie des estuaires étaient fortement impactées par un indice de contamination globale des eaux estuariennes. Cet indice de contamination globale est considéré comme un bon proxy des impacts anthropiques qui touchent les estuaires. Sur la base de ces premiers résultats, le présent travail présente une méthodologie pour la création d'indicateurs ichtyologiques liés aux masses d'eau de transition qui soient fonctionnels dans le cadre de la DCE : détermination de seuils et normes de référence permettant de distinguer entre plusieurs états écologiques de la masse d'eau, approche préliminaire pour la détermination d'un effort d'échantillonnage minimal, etc. Cette méthodologie a été développée sur l'exemple de la fonction de nourricerie des estuaires.

The extraction of oil from oilseed in an aqueous medium aims to remove the solvents used in the crushing industry. To extract oil from seeds, the enzymatic mixtures and numerous physico-chemical parameters have been studied. Two consecutive experimental designs, realized on the A.I.M. prepared from treated seeds, allowed to select effective enzyme mixtures, consisting of 3 or 4 commercial enzyme preparations out of the 25 originally selected. This original approach, based on the use of statistical tools, allows to achieve optimal mixtures for the degradation of the walls of studied seeds. The extraction of oil shows 4 phases after separation by centrifugation: solid residue, emulsion, aqueous phase and free oil. The effects of different physico-chemical parameters were evaluated (grinding time, temperature, agitation, separation...). Finally, the oil released from seeds using enzymatic mixtures made (85% of the total oil for rapeseed, 92 % for sunflower), the amount of fermentable sugars produced (60 and 80 g/l depending on conditions) or the concentration of solubilized protein (greater than 30g/l) are the landmark results of this study. This study, decrypting the conditions to implement the maximization of oil extraction in aqueous phase, is an important whole of knowledge to contribute the future development of a complete biorefining of rapeseed and sunflower with the use of each co-products : oils, proteins (napins from rape) and fermentable sugars., L'extraction d'huile de graines d'oléagineux en milieu aqueux a pour principal objectif la suppression des solvants utilisés dans les industries de trituration. Pour extraire l'huile des graines, les mélanges enzymatiques et de nombreux paramètres physico-chimiques ont été étudiés. Deux plans d'expériences consécutifs, réalisés sur le M.I.A. ont permis de formuler des mélanges d'enzymes efficaces, constitués de 3 ou 4 préparations enzymatiques commerciales parmi les 25 retenues initialement. Cette démarche permet de réaliser des mélanges optimaux pour la dégradation des parois des graines étudiées. Les extractions d'huile font apparaître 4 phases après séparation par centrifugation : résidu solide, émulsion, phase aqueuse et huile libre. Les effets de différents paramètres physico-chimiques ont été évalués (broyage, temps, température, agitation, séparation...). Au final, l'huile libérée des graines à l'aide des mélanges formulés (85% de l'huile totale pour le colza, 92% pour le tournesol), la quantité de sucres fermentescibles produite (entre 60 et 80 g/l en fonction des conditions) ou encore la concentration de protéines solubilisées (supérieure à 30g/l) sont les résultats marquants de cette étude. Cette étude décryptant les conditions à mettre en oeuvre pour maximiser l'extraction d'huile en phase aqueuse constitue un socle de connaissances permettant d'envisager le développement futur d'un bioraffinage complet des graines de colza grâce à la valorisation de chacun des co-produits huiles, protéines (napines du colza) et sucres fermentescibles.

The extraction of oil from oilseed in an aqueous medium aims to remove the solvents used in the crushing industry. To extract oil from seeds, the enzymatic mixtures and numerous physico-chemical parameters have been studied. Two consecutive experimental designs, realized on the A.I.M. prepared from treated seeds, allowed to select effective enzyme mixtures, consisting of 3 or 4 commercial enzyme preparations out of the 25 originally selected. This original approach, based on the use of statistical tools, allows to achieve optimal mixtures for the degradation of the walls of studied seeds. The extraction of oil shows 4 phases after separation by centrifugation: solid residue, emulsion, aqueous phase and free oil. The effects of different physico-chemical parameters were evaluated (grinding time, temperature, agitation, separation...). Finally, the oil released from seeds using enzymatic mixtures made (85% of the total oil for rapeseed, 92 % for sunflower), the amount of fermentable sugars produced (60 and 80 g/l depending on conditions) or the concentration of solubilized protein (greater than 30g/l) are the landmark results of this study. This study, decrypting the conditions to implement the maximization of oil extraction in aqueous phase, is an important whole of knowledge to contribute the future development of a complete biorefining of rapeseed and sunflower with the use of each co-products : oils, proteins (napins from rape) and fermentable sugars., L'extraction d'huile de graines d'oléagineux en milieu aqueux a pour principal objectif la suppression des solvants utilisés dans les industries de trituration. Pour extraire l'huile des graines, les mélanges enzymatiques et de nombreux paramètres physico-chimiques ont été étudiés. Deux plans d'expériences consécutifs, réalisés sur le M.I.A. ont permis de formuler des mélanges d'enzymes efficaces, constitués de 3 ou 4 préparations enzymatiques commerciales parmi les 25 retenues initialement. Cette démarche permet de réaliser des mélanges optimaux pour la dégradation des parois des graines étudiées. Les extractions d'huile font apparaître 4 phases après séparation par centrifugation : résidu solide, émulsion, phase aqueuse et huile libre. Les effets de différents paramètres physico-chimiques ont été évalués (broyage, temps, température, agitation, séparation...). Au final, l'huile libérée des graines à l'aide des mélanges formulés (85% de l'huile totale pour le colza, 92% pour le tournesol), la quantité de sucres fermentescibles produite (entre 60 et 80 g/l en fonction des conditions) ou encore la concentration de protéines solubilisées (supérieure à 30g/l) sont les résultats marquants de cette étude. Cette étude décryptant les conditions à mettre en oeuvre pour maximiser l'extraction d'huile en phase aqueuse constitue un socle de connaissances permettant d'envisager le développement futur d'un bioraffinage complet des graines de colza grâce à la valorisation de chacun des co-produits huiles, protéines (napines du colza) et sucres fermentescibles.

The laccase of Myceliophtora thermophila oxidizes some hydroxycinnamic acids with ephemeral formation of intermediates of different colors before these evolve spontaneously in dark brown polymers. When this oxidation is performed in biphasique medium (phosphate buffer pH 7.5 and ethyl acetate), a part of these colored intermediates can be recovered in the organic phase and it can be subtracted from an ulterior polycondensation. In the case of ferulic acid, we could isolate an orange yellow fraction, which is especially composed of dimers resulting from the condensation of the semiquinones initially formed. The synthesis of these colorants can be enhanced and their yield can be improved by controlling the reaction through a regulated minimum addition of oxygen. They keep the antioxidant power of related ferulic acid, but in high concentration (from 100 to 200 mg/ml), their cytotoxicity toward the human normal cells (HUVEC) and cancerous one (Caco-2) is important and consequently, their interest is limited as natural colorants. The semiquinone of ferulic acid doesn't form any mixed dimers, when the laccase-catalysed ferulic acid oxidation is performed in the presence of other phenolic acids. This can be explained by the fact that laccase from M. thermophila performes a multioxidation in the function of its high or little affinity for the involved substrates. In presence of an insoluble amino polyoside as chitosan, the intermediates of laccase-catalysed oxidation of different phenolic compounds (ferulic, sinapic, syringic acids and catechin) react with its NH2 groups forming covalent liaisons. Actually, this type of link leads to colored chitosans endowed of news properties due to the grafting of phenolic entities. These functionalized chitosans keep the filmogens properties of the native one. Moreover, also it can form more viscous solutions and become soluble in acidic and basic medium. It can permit the growth of HUVEC cells. They especially acquired some antioxidant properties and formed impermeable films to the oxygen which highlights multiple interesting applications, La laccase de Myceliophtora thermophila oxyde certains acides hydroxycinnamiques avec formation éphémère d'intermédiaires de différentes couleurs avant que ceux-ci n'évoluent ensuite spontanément en polymères brun noir. Quand cette oxydation est effectuée en milieu biphasique (tampon phosphate pH 7,5 / acétate d'éthyle), une partie de ces intermédiaires colorés peut être récupérée dans la phase organique et ainsi soustraite à la polycondensation ultérieure. Dans le cas de l'acide férulique, on peut ainsi isoler une fraction colorée en jaune orangé surtout composée de dimères résultant de la condensation des semiquinones initialement formées. La synthèse de ces colorants peut être favorisée et le rendement amélioré en contrôlant la réaction par un ajout minimum régulé d'oxygène. Ces colorants conservent le pouvoir antioxydant de l'acide férulique parent mais à concentration élevée (100 à 200 mg/ml), ils présentent une cytotoxicité plus élevée vis-à-vis de cellules humaines normales (HUVEC) et cancéreuses (Caco-2) ce qui limite éventuellement leur intérêt comme colorants naturels. Les semi quinones de l'acide férulique ne forment pas de dimères mixtes, quand on effectue l'oxydation laccasique de cet acide en présence d'autres acides phénoliques car la laccase de M. thermophila effectue une oxydation séquencée en fonction de sa plus ou moins grande affinité pour les différents substrats mis en jeu. En présence d'un polyoside aminé insoluble comme le chitosane, les intermédiaires d'oxydation laccasique de différents composés phénoliques (acides férulique, sinapique, syringique et catéchine) réagissent avec les groupements NH2 pour former des liaisons de covalence et conduire ainsi à des chitosanes colorés doués de nouvelles propriétés dues au greffage d'entités phénoliques. Ces chitosanes fonctionnalisés conservent les propriétés filmogènes du chitosane natif mais en plus, forment des solutions plus visqueuses et sont devenus solubles en milieux acide et basique. Ils permettent la croissance de cellules HUVEC. Ils ont surtout acquis des propriétés antioxydantes et forment des films imperméables à l'oxygène ce qui laisse entrevoir de multiples applications intéressantes

The induction of the ferulic acid esterase (FAE) synthesis was studied with Streptomyces ambofaciens ATCC 23877. The highest activity was detected in the presence of either destarched wheat bran or oat spelt xylan (0,22, 0,21 mU/mg protein, respectively). FAE productions in bioreactor were also carried out using 1% (w/v) of wheat bran as inducer. The FAE production level was three times higher in bioreactor than in Erlenmeyer flask. FAE of Streptomyces ambofaciens ATCC 23877 and that of Humicola sp., present in an enzymatic commercial mixture (DepolTM 740L), were partially purified and characterised. At the end of the purification, FAE activity of Streptomyces ambofaciens ATCC 23877 was too weak to be used later in synthesis. However, the FAE potential of Humicola sp., concentrated by acetone precipitation, for the synthesis of various phenolic esters was tested. The best conversion yields were observed in the absence of substitution on the phenolic acid aromatic cycle or in the presence of hydroxyl groups. The synthesis in non-aqueous medium (M2B2) were unsuccessful maybe because of an harmful effect of the solvent on the enzyme, L'induction de la synthèse d'une acide férulique estérase (AFE) a été étudiée chez Streptomyces ambofaciens ATCC 23877. L'activité la plus élevée a été détectée en présence de son de blé désamidonné ou de xylane d'avoine (0,22, 0,21 mU/mg protéine, respectivement). Des productions d'AFE en bioréacteur ont également été réalisées en utilisant 1% (p/v) de son de blé comme inducteur. Le niveau de production de l'AFE a été trois fois plus important en bioréacteur qu'en fiole d'Erlenmeyer. L'AFE de Streptomyces ambofaciens ATCC 23877 et celle de Humicola sp., présente dans un mélange enzymatique commercial (DepolTM 740L), ont été partiellement purifiées et caractérisées. A l'issue de la purification, l'activité AFE de Streptomyces ambofaciens ATCC 23877 a été trop faible pour pouvoir être utilisée ultérieurement en synthèse. Par contre, le potentiel de l'AFE de Humicola sp., concentrée par précipitation à l'acétone, pour la synthèse de différents esters phénoliques a été testé. Les meilleurs rendements de conversion ont été observés lors de l'absence de substitutions sur le cycle aromatique de l'acide phénolique ou en présence de groupements hydroxyles. Les synthèses en milieu non aqueux (M2B2) se sont montrées infructueuses en raison, peut-être, d'un effet néfaste du solvant sur l'enzyme