Éditeur(s) : HAL CCSD Elsevier
Résumé : Extracting and purifying a representative fraction of bacteria from soil is necessary for the application of many techniques of microbial ecology. Here the influence of different soil types on the quantitative and qualitative recovery of bacteria by soil grinding and Nycodenz density gradient centrifugation was investigated. Three soils presenting contrasted physicochemical characteristics were used for this study. For each soil, the total (AODC: acridine orange direct count) and culturable (cfu: colony-forming units) bacterial densities were measured in three distinct fractions: (i) the primary soil, (ii) the soil pellet (soil remaining after centrifugation), and (iii) the extracted cells. The automated-ribosomal intergenic spacer analysis (A-RISA) was used to characterize the community structure directly from the DNA extracted from each fraction. The physicochemical characteristics of soils were found to influence both the efficiency of bacterial cell recovery and the representativeness of the extracted cells in term of community structures between the different fractions. Surprisingly, the most representative extracted cells were obtained from the soil exhibiting the lowest efficiency of cell recovery. Our results demonstrated that quantitative and qualitative cell recovery using Nycodenz density gradient centrifugation are not necessarily related and could be differentially biased according to soil type.
ISSN: 1164-5563

hal-01031904
https://hal.archives-ouvertes.fr/hal-01031904
DOI : 10.1016/j.ejsobi.2005.08.003

Éditeur(s) : HAL CCSD Nature Publishing Group
Résumé : International audience
Forest ecosystems have integral roles in climate stability, biodiversity and economic development. Soil stewardship is essential for sustainable forest management. Organic matter (OM) removal and soil compaction are key disturbances associated with forest harvesting, but their impacts on forest ecosystems are not well understood. Because microbiological processes regulate soil ecology and biogeochemistry, microbial community structure might serve as indicator of forest ecosystem status, revealing changes in nutrient and energy flow patterns before they have irreversible effects on long-term soil productivity. We applied massively parallel pyrosequencing of over 4.6 million ribosomal marker sequences to assess the impact of OM removal and soil compaction on bacterial and fungal communities in a field experiment replicated at six forest sites in British Columbia, Canada. More than a decade after harvesting, diversity and structure of soil bacterial and fungal communities remained significantly altered by harvesting disturbances, with individual taxonomic groups responding differentially to varied levels of the disturbances. Plant symbionts, like ectomycorrhizal fungi, and saprobic taxa, such as ascomycetes and actinomycetes, were among the most sensitive to harvesting disturbances. Given their significant ecological roles in forest development, the fate of these taxa might be critical for sustainability of forest ecosystems. Although abundant bacterial populations were ubiquitous, abundant fungal populations often revealed a patchy distribution, consistent with their higher sensitivity to the examined soil disturbances. These results establish a comprehensive inventory of bacterial and fungal community composition in northern coniferous forests and demonstrate the long-term response of their structure to key disturbances associated with forest harvesting. The ISME Journal (2012) 6, 2199-2218; doi:10.1038/ismej.2012.84; published online 2 August 2012
ISSN: 1751-7362

hal-01546179
https://hal.archives-ouvertes.fr/hal-01546179
DOI : 10.1038/ismej.2012.84

Éditeur(s) : HAL CCSD
Résumé : The presence of biological matter (biofilms) in deep geological sites for storage of, for instance, toxic elements or groundwater in aquifers was clearly proved. That biomass triggers physical and chemical processes which greatly modify the durability and the sustainability of the sites. These processes, mainly from oxidative/reductive reactions, are poorly understood. This is mainly due to the fact that former studies were done at the macroscopic level far away from the micrometric scale where relevant processes induced by bacteria take place. Investigations at microscopic level are needed. Thus, we developed an experimental set-up based on the combined use of optical microscopy (transmission), atomic force microscopy (AFM) and electrical and electro-chemical AFM microscopy (EC_AFM) in order to get simultaneous information on topographic and electro-chemical processes. The first highly sensitive step was to use AFM with biological samples in liquid environment: wepresent here a study about AFM imaging of living, moving or self-immobilized bacteria, in their genuine physiological liquid medium and in true in vivo conditions. No external immobilization protocol, neither chemical nor mechanical was needed. For the first time, the native gliding movements of Gram negative Nostoc cyanobacteria upon the surface were studied by AFM. AFM height and mechanical stiffness data were simultaneously acquired. From these, mechanical parameters, inner turgor pressure and Young modulus, were derived for different bacterial species (Anabaenopsis circularis, Rhodococcus wratislaviensis). Our study revealed that AFM imaging is thus possible on moving living species. These results open a large window on new studies of both dynamical phenomena of practical and fundamental interests such as the formation of biofilms and dynamic properties of bacteria in real physiological conditions. The second delicate step was to combine AFM and optical measurements with electrical ones. We mounted a new experimental set-up coupling real-time (i) monitoring of optical properties as the optical density (OD) evolution related to bulk bacterial growth in liquid or as the counting of number of bacteria adhering on the surface of the sample as well and (ii) electrical and electrochemical measurements. Furthermore, these results will shortly be applied to the optimized monitoring of the in-situ activity of bacteria consuming oil pollutants, following this way, in real-time, the bioremediation of an oilcontaminated soil (ANR ECOTECH_BIOPHY program).
La présence de matière biologique (biofilms) dans les sites de stockage géologique profond, d'éléments toxiques ou encore de l'eau potable des aquifères est maintenant clairement démontrée. Cette biomasse est à l'origine de processus physiques et chimiques qui modifient considérablement la durabilité et la pérennité des sites concernés. Ces processus, principalement de type oxydo-réductif, sont encore mal compris. Ceci est principalement dû aux méthodes d'investigation, principalement macroscopiques, loin de l'échelle micrométrique caractéristique des bactéries. Seules des études, basées sur des méthodes d'investigation locale, peuvent apporter les informations requises. Ainsi, nous avons développé un dispositif expérimental basé sur l'utilisation combinée de la microscopie optique (en transmission), la microscopie à force atomique (AFM) et la microscopie AFM en mode électrique et électrochimique (EC_AFM) afin d'obtenir des informations simultanées sur la topographie de l'échantillon et sur les processus électrochimiques à l'échelle des bactéries. La première étape sensible consistait à utiliser l'AFM sur des échantillons biologiques en milieu liquide: nous présentons ici les résultats de l'imagerie AFM en milieu liquide de plusieurs types de bactéries dans leurs conditions physiologiques naturelles (conditions in vivo). Aucun protocole d'immobilisation, ni chimique ni mécanique, n'a été nécessaire; et pour la première fois, les mouvements de reptation de cyanobactéries Nostoc ont été étudiés par l'AFM. Les études AFM ont permis d'acquérir des données topographiques mais aussi mécaniques : nous avons pu ainsi mesurer le module d'Young, la pression de turgescence de différentes souches bactériennes (Anabaenopsis circularis, Rhodococcus wratislaviensis). Cette étude complète, a révélé que l'imagerie AFM est donc possible sur des espèces vivantes en mouvement. Ces résultats ouvrent une grande fenêtre sur de nouvelles études d'intérêts tels que la formation de biofilms et les propriétés dynamiques de bactéries dans des conditions physiologiques réelles. La deuxième étape délicate était de combiner l'AFM aux mesures optiques et électriques. Nous avons développé un nouveau dispositif expérimental permettant (i) le suivi de l'évolution de la croissance bactérienne par la mesure des propriétés optiques comme la densité optique DO (pour le développement bactérien en volume - milieu planctonique) , ou l'analyse de l'image du substrat par comptage du nombre de bactéries sur la surface de l'échantillon (biofilm), et (ii) les mesures électriques et électrochimiques. L'ensemble de ces résultats sera prochainement appliqué au développement de nouveaux outils de surveillance d'une biodépollution de terrain contaminé par les hydrocarbures, par le suivi in situ et en temps réel de l'activité de bactéries dépolluantes (ECOTECH_BIOPHY ANR).
https://tel.archives-ouvertes.fr/tel-01044714

tel-01044714
https://tel.archives-ouvertes.fr/tel-01044714
https://tel.archives-ouvertes.fr/tel-01044714/document
https://tel.archives-ouvertes.fr/tel-01044714/file/theseDhahri2013.pdf

Éditeur(s) : Université Bordeaux 1
Résumé : The mercury cycle was explored in the Petit-Saut reservoir / Sinnamary Estuary continuum in French Guiana. Main processes involve (i) photo-induced reduction, (ii) adsorption and desorption at the particle surfaces, (iii) methylating bacterial activity, (iv) precipitation and dissolution of iron oxyhydroxides and sulfides. Sources of mercury to the Petit-Saut reservoir consist of the partly impacted river-tributaries, the atmospheric deposition and the flooded soils or vegetation. Once into the reservoir, divalent mercury undergoes a large spectrum of transformations including speciation and phase changes. We, here, probe these processes and demonstrate that the Petit-Saut reservoir behaves as a privileged site for atmospheric recycling and in situ methylation. At the air / water interface, the mercury evasion exceeds by 50 % the direct atmospheric deposition. The in situ monomethylmercury (MMHg) production mostly occurs at the water-column chemocline and near the benthic interface. It reaches, for the whole reservoir, 8.1 moles yr-1 which corresponds to a 0.06 % d-1 methylation rate. We calculated that the 2003/04 period MMHg outputs from the reservoir to the Sinnamary Estuary averaged 13.5 moles yr-1 while the inputs were 5.4 moles yr-1. The dynamic Sinnamary Estuary is the genuine geochemical reactor of the continuum. There, the divalent mercury undergoes rapid yet profound partitioning among phases, recycling to the atmosphere and methylation processes. Those mostly originate from the long-scale oxygen paucity of the dam discharged waters. The expression of marked redox gradients and the intense organic matter turnover enhance the production of methylable inorganic mercurial species and tend to favor the sulfate reducing bacteria (methylating agents) activity. As a result, the dynamic Sinnamary Estuary exports 60 % more MMHg than it does import: 27 versus 17 moles yr-1. Further at large, in the saline estuary, we determined: (i) a non conservative behavior of dissolved (< 0.2 µm) mercury, (ii) a lowering of the mercury level of particles during mixing with the Amazon plume and (iii) a gradual dilution of the estuarine mercury concentrations by oceanic waters.
La retenue de Petit-Saut et, à l'aval, l'estuaire du Sinnamary sont deux centres d'altération du cycle naturel du mercure. Les processus sont, entre autres, (i) la photoréduction, (ii) la sorption / désorption en surface des particules, (iii) l'activité méthylante des bactéries, (iv) la dissolution / précipitation des oxyhydroxydes et des sulfures de fer. Les sources de mercure vers la retenue sont variées. Elles regroupent, en premier lieu, les affluents (partiellement orpaillés), l'atmosphère, les sols et la végétation ennoyés. Dans la colonne d'eau, les formes divalentes du mercure subissent un train complet de transformations couplant changements de phase et de spéciation. Nos résultats illustrent ces processus et démontrent que la retenue constitue un site privilégié de recyclage vers l'atmosphère et de méthylation. A l'interface air / eau, les exports en mercure de la retenue excèdent de 50 % le dépôt atmosphérique. La production endogène de monométhylmercure (MMHg) a lieu dans des sites particuliers : la chemocline et l'interface benthique. Elle atteint pour l'ensemble de la retenue 8,1 moles a-1 soit un taux de méthylation de 0,06 % j-1. On estime que la retenue exporte vers l'estuaire du Sinnamary 13,5 moles a-1 de MMHg alors qu'elle n'importe que 5,4 moles a-1. L'estuaire dynamique du Sinnamary constitue le véritable réacteur chimique du système fonctionnant en liaison avec celui de la retenue. Le mercure y décrit un cycle rapide aux processus de partition, de recyclage et de méthylation marqués. Ces processus résultent de l'oxygénation incomplète et durable des masses d'eau anoxiques exportées par la retenue de Petit-Saut. L'expression de gradients rédox prononcés conjuguée à un intense recyclage de la matière organique y favorise la production de formes méthylables de mercure ainsi que l'activité des bactéries sulfatoréductrices (agents de la méthylation). C'est pourquoi, l'estuaire dynamique du Sinnamary exporte 60 % plus de MMHg qu'il n'importe : 27 contre 17 moles a-1. Plus en aval, dans l'estuaire salin, nous avons mis en évidence : (i) un comportement non conservatif du mercure dissous, (ii) un appauvrissement du mercure des particules suite au mélange avec le panache de l'Amazone et (iii) une dilution des concentrations par les masses d'eau océaniques.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2006/these-1508.pdf
http://archimer.ifremer.fr/doc/00000/1508/

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
Banana has been a main agricultural product in the French West Indies (Guadeloupe and Martinique) since the 1960s. This crop requires the intensive use of pesticides to prevent attacks by insect pests. Chlorinated pesticides, such as hexachlorocyclohexane (HCH), chlordecone and dieldrin, were used until the beginning of the 1990s, resulting in a generalized diffuse contamination of the soil and water in the areas of banana production, hence the need to develop solutions for cleanup of the polluted sites. The aims of this work were (i) to assess lindane degradation in soil slurry microcosms treated with lindane at 10 mg/L and (ii) to detect the catabolic genes involved in the HCH degradation pathway. The soil slurry microcosm system showed a 40 % lindane degradation efficiency at the end of a 30-day experiment. Lower lindane removal was also detected in the abiotic controls, probably caused by pesticide adsorption to soil particles. Indeed, the lindane concentration decreased from 6000 to 1330 ng/mL and from 800 to 340 ng/mL for the biotic and abiotic soils, respectively. Nevertheless, some of the genes involved in the HCH degradation pathway were amplified by polymerase chain reaction (PCR) from crude deoxyribonucleic acid (DNA) extracted from the Guadeloupe agricultural soil, suggesting that HCH degradation is probably mediated by bacteria closely related to the family Sphingomonadaceae.
ISSN: 0944-1344

hal-01545905
https://hal.archives-ouvertes.fr/hal-01545905
DOI : 10.1007/s11356-015-5875-7

Éditeur(s) : ISTPM
Résumé : Table of Contents The Natural Environment Production centres Oyster farming centres Mussel farming centres Chapter I. - The Physicochemical Environment I. Sea Water a) Transparency, turbidity b) Temperature c) Salinity d) Dissolved gases e) Substances dissolved in the water f) Water movements g) Hydrology of the main shellfish farming centres II. Soils a) Soil properties b) Mechanical amendment of soils, its effects c) Fertilisation of soils in shellfish farming d) Changes in soils due to farming Bibliography Chapter II. - The Natural Environment Phytoplankton Zooplankton Basic biology of phytoplankton Assessment of phytoplankton production, notion of primary productivity Phytoplankton from a few shellfish farming regions Microphytobenthos Factors controlling primary productivity Toxicity of planktonic organisms; red tides Bibliography Chapter III – Alterations in the Natural Environment Pollution I. Chemical Pollution Definitions Influence of turbidity A. Telluric pollution 1o Domestic pollution 2o Agriculture-related pollution 3o Industrial pollution B. Pelagic pollution 1o Pollution through hydrocarbons Conclusion II. Bacterial Pollution Future of micro-organisms in the sea A. Specimen conditions B. Phenomena of dispersion Survival of germs in the marine environment A. Factors influencing the total microbial content B. Resistance of various groups of germs Consequences of bacterial pollutions on shellfish and shellfish farming Shellfish Shellfish farming Conclusion Bibliography
Sommaire : Le milieu naturel Les centres de production Les centres ostréicoles Les centres mytilicoles Chapitre I. - le milieu physico-chimique I. L'eau de mer a) Transparence, turbidité b) Température c) Salinité d ) Les gaz dissous e) Les substances dissoutes dans l'eau f) Les mouvements de l'eau g) Hydrologie des principaux centres conchylicoles IL Les sols a ) Propriétés des sols b) Amendement mécanique des sols, ses effets c) La fertilisation des sols en conchylicutture d) Evolution des sols du fait de la culture Bibliographie Chapitre II. - le milieu biologique Le phytoplancton Le zooplancton Biologie sommaire du phytoplancton Evaluation de la production de phytoplancton, notion de productivité primaire Phytoplancton de quelques régions conchylicoles Le microphytobenthos Facteurs contrôlant la productivité primaire Toxicité d'organismes planctoniques ; les eaux rouges bibliographie Chapitre III - les altérations du milieu naturel. Les pollutions I. Les pollutions chimiques Définitions Influence de la turbidité A. Pollution tellurique 1° Pollution d'origine domestique 2° Pollutions liées aux activités agricoles 3° Pollutions industrielles B. Pollution pélagique 1° Pollution par les hydrocarbures Conclusion II. Les pollutions bactériennes Devenir des microorganismes dans la mer A. Conditions de prélèvements B. Phénomènes de dispersion Survie des germes dans le milieu marin A. Facteurs influant sur la teneur microbienne globale B. Résistance des divers groupes de germes Conséquences des pollutions bactériennes sur les coquillages et sur la conchyliculture Les coquillages La conchyliculture Conclusion Bibliographie
Revue des Travaux de l'Institut des Pêches Maritimes (0035-2276) (ISTPM), 1974-09 , Vol. 38 , N. 3 , P. 217-337

Droits : Ifremer
http://archimer.ifremer.fr/doc/1974/publication-1776.pdf
http://archimer.ifremer.fr/doc/00000/1776/

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
Three soil DNA extraction procedures (homemade protocols and commercial kit) varying in their practicability were applied to contrasting soils to evaluate their efficiency in recovering: (i) soil DNA and (ii) bacterial diversity estimated by 16S rDNA pyrosequencing. Significant differences in DNA yield were systematically observed between tested procedures. For certain soils, 10 times more DNA was recovered with one protocol than with the others. About 15 000 sequences of 16S rDNA were obtained for each sample which were clustered to draw rarefaction curves. These curves, as well as the PCA ordination of community composition based on OTU clustering, did not reveal any significant difference between procedures. Nevertheless, significant differences between procedures were highlighted by the taxonomic identification of sequences obtained at the phylum to genus levels. Depending on the soil, differences in the number of genera detected ranged from 1% to 26% between the most and least efficient procedures, mainly due to a poorer capacity to recover populations belonging to Actinobacteria, Firmicutes or Crenarchaeota. This study enabled us to rank the relative efficiencies of protocols for their recovery of soil molecular microbial biomass and bacterial diversity and to help choosing an appropriate soil DNA extraction procedure adapted to novel sequencing technologies.
ISSN: 1751-7915

hal-01546177
https://hal.archives-ouvertes.fr/hal-01546177
DOI : 10.1111/j.1751-7915.2011.00307.x

Éditeur(s) : Pergamon-elsevier Science Ltd
Résumé : Total mercury (HgT) and monomethylmercury (MMHg) were investigated in a tropical head watershed (1 km(2)) of French Guiana. The watershed includes a pristine area on the hill slopes and a former gold mined flat in the bottomland. Concentrations of dissolved and particulate HgT and MMHg were measured in rain, throughfall, soil water and at three points along the stream. Samples were taken in-between and during 14 storm events at the beginning and middle of the 2005 and 2006 rainy seasons. Dissolved and particulate HgT concentrations in the stream slightly increased downstream, while dissolved and particulate MMHg concentrations were low at the pristine sub-watershed outlet (median = 0.006 ng L-1 and 1.84 ng g(-1), respectively) and sharply increased at the gold mined flat outlet (median = 0.056 ng L-1 and 6.80 ng g(-1), respectively). Oxisols, which are dominant in the pristine area act as a sink of HgT and MMHg from rain and throughfall inputs. Hydromorphic soils in the flat are strongly contaminated with Hg (including Hg droplets) and their structure has been disturbed by former gold-mining processes, leading to multiple stagnant water areas where biogeochemical conditions are favorable for methylation. In the former gold mined flat high dissolved MMHg concentrations (up to 0.8 ng L-1) were measured in puddles or suboxic soil pore waters, whereas high dissolved HgT concentrations were found in lower Eh conditions. Iron-reducing bacteria were suggested as the main methylators since highest concentrations for dissolved MMHg were associated with high dissolved ferrous iron concentrations. The connection between saturated areas and stagnant waters with the hydrographic network during rain events leads to the export of dissolved MMHg and HgT in stream waters, especially at the beginning of the rainy season. As both legal and illegal gold-mining continues to expand in French Guiana, an increase in dissolved and particulate MMHg emissions in the hydrographic network is expected. This will enhance MMHg bio-amplification and present a threat to local populations, whose diet relies mainly on fish.
Water Research (0043-1354) (Pergamon-elsevier Science Ltd), 2011-04 , Vol. 45 , N. 8 , P. 2659-2669

Droits : 2011 Elsevier Ltd All rights reserved.
http://archimer.ifremer.fr/doc/00035/14628/12134.pdf
DOI:10.1016/j.watres.2011.02.022
http://archimer.ifremer.fr/doc/00035/14628/

Éditeur(s) : HAL CCSD Wiley
Résumé : 1. Resource control over abundance, structure and functional diversity of soil microbial communities is a key determinant of soil processes and related ecosystem functioning. Copiotrophic organisms tend to be found in environments which are rich in nutrients, particularly carbon, in contrast to oligotrophs, which survive in much lower carbon concentrations. We hypothesized that microbial biomass, activity and community structure in nutrient-poor soils of an Amazonian rain forest are limited by multiple elements in interaction. We tested this hypothesis with a fertilization experiment by adding C (as cellulose), N (as urea) and P (as phosphate) in all possible combinations to a total of 40 plots of an undisturbed tropical forest in French Guiana. After 2years of fertilization, we measured a 47% higher biomass, a 21% increase in substrate-induced respiration rate and a 5-fold higher rate of decomposition of cellulose paper discs of soil microbial communities that grew in P-fertilized plots compared to plots without P fertilization. These responses were amplified with a simultaneous C fertilization suggesting P and C colimitation of soil micro-organisms at our study site. Moreover, P fertilization modified microbial community structure (PLFAs) to a more copiotrophic bacterial community indicated by a significant decrease in the Gram-positive:Gram-negative ratio. The Fungi:Bacteria ratio increased in N fertilized plots, suggesting that fungi are relatively more limited by N than bacteria. Changes in microbial community structure did not affect rates of general processes such as glucose mineralization and cellulose paper decomposition. In contrast, community level physiological profiles under P fertilization combined with either C or N fertilization or both differed strongly from all other treatments, indicating functionally different microbial communities. While P appears to be the most critical from the three major elements we manipulated, the strongest effects were observed in combination with either supplementary C or N addition in support of multiple element control on soil microbial functioning and community structure. We conclude that the soil microbial community in the studied tropical rain forest and the processes it drives is finely tuned by the relative availability in C, N and P. Any shifts in the relative abundance of these key elements may affect spatial and temporal heterogeneity in microbial community structure, their associated functions and the dynamics of C and nutrients in tropical ecosystems.
ISSN: 0269-8463

hal-01204229
https://hal.archives-ouvertes.fr/hal-01204229
DOI : 10.1111/1365-2435.12329
PRODINRA : 311389

Éditeur(s) : HAL CCSD
Résumé :   Diplôme : DEA
il s'agit d'un type de produit dont les métadonnées ne correspondent pas aux métadonnées attendues dans les autres types de produit : DISSERTATION
As part of a programme of degraded soils rehabilitation, and in order to follow the biological processes connected with the soils reconstitution, it is a requisite to determine indicators able to account for the evolution of the soil. These indicators can he found on structural and/or functional characteristics of the bacterial communities, such as soil respiration, denitrification or genetic diversity. Theses descriptors are able to characterize the global community or targeted communities chosen for their importance in the ecosystem functioning. In this work, we tried to know whether soil bacteria genetic diversity was able to be a relevant indicator of the soil evolution in a rehabilitated sequence. This diversity is apprehended via a molecular technique: ARISA. A chronosequence situated on a gold mine, already experimented as part of a programme of degraded zones rehabilitation, was at our disposal. Results show that ARISA is a very sensitive technique, able to account for the bacterial community heterogeneity in a soil but unable, at the parcel scale, to discriminate the treatments nor to account for the evolution in lime of planted treatments. Lastly, the advantage of this indicator of genetic diversity may arise from its confrontation to other indicators (functional, for example).
Dans le cadre de la revégétalisation d'un sol dégradé et afin de pouvoir suivre les processus biologiques liés à la reconstitution des sols, il est nécessaire de déterminer des indicateurs pouvant rendre compte de l'évolution de l'état du sol. Ces indicateurs peuvent être fondés sur des caractéristiques fonctionnelles et/ou structurales des communautés bactériennes telles que la respiration, la dénitrification ou la diversité génétique. Ces descripteurs peuvent analyser la communauté globale ou des communautés cibles choisies pour leur importance dans le fonctionnement de l'écosystème. L'objectif de ce stage était de savoir si la diversité génétique des bactéries du sol pouvait être un indicateur de l'évolution des sols au sein d'une séquence de revégétalisation. Cette diversité est appréhendée par une technique de biologie moléculaire : l'ARISA. On dispose d'une chronoséquence située sur une mine d'or étudiée dans le cadre d'un programme de réhabilitation des zones dégradées. Les résultats montrent que l'ARISA est une technique très sensible, qui rend bien compte de l’hétérogénéité des communautés bactériennes présentes dans un sol mais est incapable, à l’échelle de la parcelle, de discriminer ni les traitements ni de rendre compte de l'évolution dans le temps de ces parcelles. Enfin, l'intérêt de cet indicateur de diversité génétique pourra résulter de sa confrontation avec d'autres indicateurs (fonctionnels par exemple).
https://hal.archives-ouvertes.fr/hal-01189155

hal-01189155
https://hal.archives-ouvertes.fr/hal-01189155
PRODINRA : 9464

Éditeur(s) : HAL CCSD Elsevier
Résumé : DNA metabarcoding on soil samples is increasingly used for large-scale and multi-taxa biodiversity studies. However, DNA extraction may be a major bottleneck for such wide uses. It should be cost/time effective and allow dealing with large sample volumes so as to maximise the representativeness of both micro- and macro-organisms diversity. Here, we compared the performances of a fast and cheap extracellular DNA extraction protocol with a total DNA extraction method in retrieving bacterial, eukaryotic and plant diversity from tropical soil samples of ca. 10 g. The total DNA extraction protocol yielded more high-quality DNA. Yet, the extracellular DNA protocol provided similar diversity assessments although it presented some differences in clades relative abundance and undersampling biases. We argue that extracellular DNA is a good compromise between cost, labor, and accuracy for high throughput DNA metabarcoding studies of soil biodiversity.
ISSN: 0038-0717

hal-01532638
https://hal.archives-ouvertes.fr/hal-01532638
DOI : 10.1016/j.soilbio.2016.01.008
PRODINRA : 384522

Résumé : Les biofilms électroactifs (EA) sont des consortia bactériens mono ou pluri-espèces, qui ont la faculté d’échanger des électrons provenant de leur métabolisme avec les surfaces solides conductrices des électrodes. Cette découverte est à l’origine d’un nouveau dispositif énergétique : la pile à combustible microbienne (PACM). Depuis les années 2000, la littérature scientifique sur les biofilms EA et sur les PACM explose, notamment grâce à la découverte de bactéries capables de transférer par voie directe – par des pili ou protéines transmembranaires – des électrons vers les électrodes. Plusieurs sources de bactéries EA ont été à ce jour découvertes, allant de cultures pures à des communautés bactériennes plus complexes. Ces dernières sont issues de milieux aqueux naturels (milieux marins ou d’eau douce), industriels ou urbains (effluents d’industrie, eaux usées domestiques). La plupart de ces sources bactériennes proviennent d’environnements de climat tempéré. Dans ce travail de thèse, plusieurs sols de milieux humides de la Guyane ont été identifiés comme étant de sources de bactéries EA. Les expériences menées sous potentiel d’électrode imposé et constant ont permis d’étudier l’adhésion à la surface d’électrode des biofilms EA issus de la flore endogène des milieux sélectionnés. La formation de bioanodes et de biocathodes a été possible en présence respectivement d’acétate et d’oxygène dans les milieux. Une étude par voltammétrie cyclique a mis en évidence les pics d’oxydo-réduction en lien avec les échanges électroniques du biofilm EA et de l’électrode. En optimisant la procédure de formation des biofilms EA par chronoampérométrie (surface d’électrode plus importante, apport en continu et progressif du substrat), une densité de courant maximale de 12 A/m2 et un rendement faradique de 24 % ont été obtenus. Une autre méthode pour former des biofilms EA à partir d’un milieu choisi, la mangrove, a consisté à utiliser deux prototypes de PACM : une pile à compartiment unique et à cathode à air, et une pile benthique. Dans les deux cas, les biofilms EA ont pu être formés et étudiés, complétant certaines observations faites sous potentiel imposé. La PACM benthique s’est avérée être la plus proche d’une application à grande échelle puisqu’elle a été complètement autonome : anode et cathode utilisant uniquement le milieu pour fonctionner. Elle a pu être étudiée en laboratoire comme sur le terrain.
Electroactive biofilms (EA) are mono or multi-species bacterial consortia, which have the ability to exchange electrons from their metabolism with solid surfaces of conductive electrodes. This discovery is the basis for a device of energy production: microbial fuel cell (MFC). Since the 2000s, the scientific literature on EA biofilms and MFC explodes, thanks to the discovery of bacteria that are able to transfer directly – by pili or trans-membrane proteins – electrons to electrodes. Several sources of EA bacteria were discovered to date, ranging from pure cultures to more complex bacterial communities. Those last are from natural (marine and freshwater), industrial or urban (industrial effluents, domestic wastewater) aqueous environments. However, the vast majority of these are from temperate environments.In this thesis, several wetland soils of French Guiana have been identified as sources of EA bacteria. Experiments under poised and constant electrode potential were used to examine adherence to the electrode surface of EA biofilms from the endogenous flora of selected environments. Formation of bioanodes and biocathodes was possible in the presence respectively of acetate and oxygen in the media. A study by cyclic voltammetry showed the redox peaks related to electronic exchanges between EA biofilm and electrode. By optimizing the process of EA biofilm formation by chronoamperometry (larger electrode surface, providing continuous and progressive substrate), a maximum current density of 12 A/m2 and a coulombic efficiency of 24% were obtained.Another method to form EA biofilms from a chosen medium (mangrove) was to use two MFC prototypes: a single compartment and air cathode one, and a benthic one. In both cases, the EA biofilms have been trained and studied supplementing certain observations made under poised polarization. MFC benthic proved to be the closest to a wide application since it was completely autonomous, anode and cathode only using the medium to function. It has been studied in the laboratory and in the field.

http://www.theses.fr/2012AGUY0560/document

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : Functional aspects of biodiversity were investigated in a lowland tropical rainforest in French Guyana (5°2'N, annual precipitation 2200 mm). We assessed leaf '15N as a presumptive indicator of symbiotic N2 fixation, and leaf and wood cellulose '13C as an indicator of leaf intrinsic water-use efficiency (CO2 assimilation rate/leaf conductance for water vapour) in dominant trees of 21 species selected for their representativeness in the forest cover, their ecological strategy (pioneers or late successional stage species, shade tolerance) or their potential ability for N2 fixation. Similar measurements were made in trees of native species growing in a nearby plantation after severe perturbation (clear cutting, mechanical soil disturbance). Bulk soil '15N was spatially quite uniform in the forest (range 3-5‰), whereas average leaf '15N ranged from m0.3‰ to 3.5‰ in the different species. Three species only, Diplotropis purpurea, Recordoxylon speciosum (Fabaceae), and Sclerolobium melinonii (Caesalpiniaceae), had root bacterial nodules, which was also associated with leaf N concentrations higher than 20 mg gm1. Although nodulated trees displayed significantly lower leaf '15N values than non-nodulated trees, leaf '15N did not prove a straightforward indicator of symbiotic fixation, since there was a clear overlap of '15N values for nodulated and non-nodulated species at the lower end of the '15N range. Perturbation did not markedly affect the difference '15Nsoil m '15Nleaf, and thus the isotopic data provide no evidence of an alteration in the different N acquisition patterns. Extremely large interspecific differences in sunlit leaf '13C were observed in the forest (average values from m31.4 to m26.7‰), corresponding to intrinsic water-use efficiencies (ratio CO2 assimilation rate/leaf conductance for water vapour) varying over a threefold range. Wood cellulose '13C was positively related to total leaf '13C, the former values being 2-3‰ higher than the latter ones. Leaf '13C was not related to leaf '15N at either intraspecific or interspecific levels. '13C of sunlit leaves was highest in shade hemitolerant emergent species and was lower in heliophilic, but also in shade-tolerant species. For a given species, leaf '13C did not differ between the pristine forest and the disturbed plantation conditions. Our results are not in accord with the concept of existence of functional types of species characterized by common suites of traits underlying niche differentiation; rather, they support the hypothesis that each trait leads to a separate grouping of species.
ISSN: 0029-8549

hal-01032378
https://hal.archives-ouvertes.fr/hal-01032378
DOI : 10.1007/s004420050593

Éditeur(s) : HAL CCSD
Résumé : Background and objectives.[br/] Dairy propionibacteria are isolated from various ecological niches, including soil, grass, silage, rumen, raw milk and dairy plants, showing great adaptability and robustness. Used as ripening starters in Swiss type cheeses, these food grade bacteria are described as nutraceutical producers and they release into the external medium short chain fatty acids (SCFA), folic acid, cobalamin and the bifidogenic 1,4- dihydroxy-2-naphthoic acid (DHNA). These compounds are known to play a pivotal role in the modulation of intestinal physiology through diet. Considering the pro-apoptotic potential of some of these metabolites, we investigated the potential of dairy products fermented by the main species, Propionibacterium freudenreichii, to prevent colon carcinogenesis. Methods.[br/] We developed a new fermented milk. Taking advantage of milk fractions, high and stable populations of P. freudenreichii were obtained in this product. It was tested in vitro on gastric HGT-1 and colic HT-29 cells with respect to the typical hallmarks of apoptosis. In vivo, the fermented milk was evaluated in piglets with respect to general health parameters and in human microbiota-associated rats, treated or not with dimethylhydrazine (DMH), with respect to intestinal proliferation and apoptosis. Results. Fermented milk induced cell death in both gastric and colon cancer cells. Cellular and molecular characterization of cells dying in response to SCFA treatment revealed cell cycle arrest, drop in intracellular ATP, depolarization of mitochondria, translocation of key apoptosis proteins, processing of caspases and fragmentation of the nucleus. Interestingly, the new fermented milk was shown to potentiate the cytotoxic effect of molecules used in cancer chemotherapy. In accordance, it modulated expression of key apoptotic proteins. In vivo, the fermented milk was evaluated in piglets. Daily oral gavage led to high colic P. freudenreichii populations, to modulation of the intestinal microbiota and modulation of intestinal cytokines. Promotion of piglets’ growth also revealed general probiotic effects of fermented milk. Considering that propionibacteria were metabolically active in the intestine, we further investigated their impact on colon carcinogenesis (induced by dimethylhydrazine, DMH) in human microbiota-associated rats. Consumption of selected strains of propionibacteria resulted in enhanced apoptotic depletion of DMH damaged epithelial cells, yet had no effect on neither proliferation nor apoptosis in healthy rats. Conclusions and perspectives.[br/] These results open new perspectives in the field of colon cancer cells prevention and/or treatment. The synergy with pro-apoptotic chemotherapy molecules suggests that such a fermented product may be proposed as a food supplement to enhance the effects of anticancer treatments.
colloque international « Nutrition and Cancer : Hot topics from biology to public health issues »
Paris, France

hal-01190452
https://hal.archives-ouvertes.fr/hal-01190452
PRODINRA : 201853

Éditeur(s) : Wiley-blackwell Publishing, Inc
Résumé : Aims: A description of bacterial pathogens in shrimp ponds is necessary to understand their pathogenesis. Vibrio nigripulchritudo was shown to contain saprophytic and pathogenic strains among New Caledonian isolates. We established a method to map the development of V. nigripulchritudo in pond sediments at three different genetic levels: the species level, then at the pathogenic cluster level and finally at the plasmid level, present only in all highly pathogenic isolates. Methods and Results: PCR methods were applied to shrimp pond sediments both before and after a mortality outbreak. Using crude samples, the species V. nigripulchritudo is not detected at first (0/42 samples at day 56 post stocking) but appears frequently in the sediments after the mortality event (30/42 at day 107). The distribution of strains from the pathogenic cluster of V. nigripulchritudo also follows this pattern. In contrast, the pSFn1 virulence-associated plasmid was detected in one sample at day 56 and none at day 107. An enrichment method was developed to lower the detection limits of our assays. After enrichment, the species V. nigripulchritudo was detected in all samples at both dates. The number of samples positive for pSFn1 was 42/42 samples at day 56 and 29/42 at day 107. Conclusions: These results show that the sediments contain V. nigripulchritudo, notably pathogenic strains. Surprisingly, the virulence-associated plasmid pSFn1 found in all V. nigripulchritudo isolated from moribund shrimp appears less frequently in sediments, possibly being useless or even detrimental to its recipient bacteria in this environment. Significance and Impact of the Study: This study confirms the presence of pathogenic V. nigripulchritudo strains in shrimp pond sediment before a mortality outbreak complying with a previous hypothesis that sediments could be the infecting reservoir. After the outbreak, both total V. nigripulchritudo and pathogenic strains populations have largely increased, possibly contributing to the recurrent mortality observed in this shrimp vibriosis.
Journal Of Applied Microbiology (1364-5072) (Wiley-blackwell Publishing, Inc), 2010-06 , Vol. 108 , N. 6 , P. 2003-2011

Droits : 2010 The Society for Applied Microbiology - Wiley-Blackwell
http://archimer.ifremer.fr/doc/00004/11484/8177.pdf
DOI:10.1111/j.1365-2672.2009.04601.x
http://archimer.ifremer.fr/doc/00004/11484/

Résumé : L’ingénierie agroécologique vise à produire des savoirs actionnables, pour concevoir des systèmes de cultures économiquement et écologiquement performants, par la valorisation de régulations naturelles. Notre problématique est centrée sur la santé des cultures, et plus particulièrement sur la bactérie phytopathogène Ralstonia solanacearum, agent du flétrissement bactérien alors qu’une souche extrêmement agressive menace la production de tomates en plein champ en Martinique. La nécessité d’explorer et de développer des alternatives aux méthodes conventionnelles de protection des plantes (variétés résistantes, pesticides), actuellement inefficaces, invite à la mise en œuvre d’une démarche de conception innovante. Nos travaux montrent que la mobilisation d’une barrière rhizosphérique est une stratégie de régulation biologique alternative. Différents processus y contribuent, telle que la mycorhization, symbiose entre racines et champignons mycorhiziens à arbuscules, présents dans la plupart des sols. Nous montrons que la mobilisation de réseaux de mycorhizes indigènes à partir d’un sol agricole permet une mycorhization précoce de la tomate. De plus, l’association de plantes aux propriétés mycorhizotrophes et assainissantes en conditions contrôlées montre des effets bioprotecteurs partiels et ouvre de nouvelles perspectives de combinaisons entre processus. Ces combinaisons sont mobilisables par des leviers d’actions multi-scalaires. Nous avons produit une grille d’analyse générique de ces leviers d’action pour la conception, par des trajectoires d’innovation multidirectionnelles, de « systèmes de culture bioprotégés ». Dans le contexte agricole martiniquais, une démarche d’apprentissage permet en effet l’émergence d’une dynamique de co-conception de systèmes de cultures recourant aux plantes mycorhizotrophes. Nos travaux proposent des outils pour une exploration collective de nouvelles stratégies de gestion durable de la santé des cultures.
Agroecological engineering aims to produce actionable knowledge to design economically and environmentally efficient cropping systems, based on the exploitation of natural regulation mechanisms. Our issue is centered on crop health, especially on the plant pathogenic bacterium Ralstonia solanacearum (bacterial wilt agent), of which an extremely aggressive strain threatens field tomato production in Martinique. The need to explore and develop alternatives to conventional methods of plant protection (resistant varieties, pesticides), ineffective in our case, calls for the implementation of an innovative design approach. Our work shows that the protection of the roots via the formation of a self-sustaining rhizospheric barrier may be an alternative biological control strategy. Different processes contribute, such as mycorrhizal symbiosis between roots and arbuscular mycorrhizal fungi, which are present in most soils. We show that the mobilization of indigenous mycorrhizal networks from an agricultural soil allows early mycorrhization of tomatoes. In addition, the association of plants with mycorrhizal and sanitizing properties in controlled conditions showed partial bioprotective effects and opens up new prospects for combinations between processes. These combinations may be exploited in various ways. We produced a generic analysis grid of key levers to design "healthy cropping systems " through multi-directional innovation trajectories. In Martinique's agricultural context, a learning process allows the emergence of a dynamic co-design of cropping systems using mycorrhizal plants. Our work thus provides tools for collective exploration of new sustainable management strategies for crop health.

http://www.theses.fr/2015AGUY0828/document