Characterization of specificity of bacterial community structure within the burrow environment of the marine polychaete Hediste (Nereis) diversicolor

Bioturbation is known to stimulate microbial communities, especially in macrofaunal burrows where the abundance and activities of bacteria are increased. Until now, these microbial communities have been poorly characterized and an important ecological question remains: do burrow walls harbor similar or specific communities compared with anoxic and surface sediments? The bacterial community structure of coastal sediments inhabited by the polychaete worm Hediste diversicolor was investigated. Surface, burrow wall and anoxic sediments were collected at the Carteau beach (Gulf of Fos, Mediterranean Sea). Bacterial diversity was determined by analyzing small subunit ribosomal RNA (16S rRNA) sequences from three clone libraries (168, 179 and 129 sequences for the surface, burrow wall and anoxic sediments, respectively). Libraries revealed 306 different operational taxonomic units (OTUs) belonging to at least 15 bacterial phyla. Bioinformatic analyses and comparisons between the three clone libraries showed that the burrow walls harbored a specific bacterial community structure which differed from the surface and anoxic environments. More similarities were nevertheless found with the surface assemblage. Inside the burrow walls, the bacterial community was characterized by high biodiversity, which probably results from the biogeochemical heterogeneity of the burrow system

PRT: Parallel program for a full backtranslation of oligopeptides

DNA hybridization methods have become the most widely used tools in molecular biology to identify organisms and evaluate gene expression levels. PCR (Polymerase Chain Reaction)-based methods, fluorescent in situ hybridization (FISH) and the recent development of DNA microarrays as a high throughput technology need efficient primers or probes design. Evaluation of the metabolic capacities of complex microbial communities found in terrestrial or aquatic environments requires new probe design algorithms that reflect the genetic diversity. As only a small part of the microbial diversity is known, gene sequences deposited in international databases do not reflect the entire diversity. In this context we propose to use oligopeptide sequences for the design of complete set of DNA probes that are able to target the entire genetic diversity of genes encoding enzymes. Due to the degenerated genetic code backtranslation must be managed efficiently. To our knowledge no software has been developed to propose a full backtranslation. This complexity is tractable since we only need to focus on short oligopeptides for DNA probe design. We propose new algorithms that perform a high performance oligopeptide backtranslation into all potential nucleic sequences. We use different efficient techniques such as memory mapping to perform such a computing. We also propose a MPI parallel computing that reduces the whole execution time using data load balancing and network file stream distribution on a cluster architecture

Dynamics of bacterial assemblages and removal of polycyclic aromatic hydrocarbons in oil-contaminated coastal marine sediments subjected to contrasted oxygen regimes

To study the impact of oxygen regimes on the removal of polycylic aromatic hydrocarbons (PAHs) in oil-spill-affected coastal marine sediments, we used a thin-layer incubation method to ensure that the incubated sediment was fully oxic, anoxic, or was influenced by oxic-anoxic switches without sediment stirring. Hydrocarbon content and microbial assemblages were followed during 60 days to determine PAH degradation kinetics and microbial community dynamics according to the oxygenation regimes. The highest PAH removal, with 69 % reduction, was obtained at the end of the experiment under oxic conditions, whereas weaker removals were obtained under oscillating and anoxic conditions (18 and 12 %, respectively). Bacterial community structure during the experiment was determined using a dual 16S rRNA genes/16S rRNA transcripts approach, allowing the characterization of metabolically active bacteria responsible for the functioning of the bacterial community in the contaminated sediment. The shift of the metabolically active bacterial communities showed that the selection of first responders belonged to Pseudomonas spp. and Labrenzia sp. and included an unidentified Deltaproteobacteria—irrespective of the oxygen regime—followed by the selection of late responders adapted to the oxygen regime. A novel unaffiliated phylotype (B38) was highly active during the last stage of the experiment, at which time, the low-molecular-weight (LMW) PAH biodegradation rates were significant for permanent oxic- and oxygen-oscillating conditions, suggesting that this novel phylotype plays an active role during the restoration phase of the studied ecosystem

Oil spill effects on macrofaunal communities and bioturbation of pristine marine sediments (Caleta Valdés, Patagonia, Argentina): experimental evidence of low resistance capacities of benthic systems without history of pollution

The Patagonian coast is characterized by the existence of pristine ecosystems which may be particularly sensitive to oil contamination. In this study, a simulated oil spill at acute and chronic input levels was carried out to assess the effects of contamination on the macrobenthic community structure and the bioturbation activity of sediments sampled in Caleta Valdés creek. Superficial sediments were either noncontaminated or contaminated by Escalante crude oil and incubated in the laboratory for 30 days. Oil contamination induced adverse effects on macrobenthic community at both concentrations with, for the highest concentration, a marked decrease of approximately 40 and 55%of density and specific richness, respectively. Besides the disappearance of sensitive species, some other species like Oligochaeta sp. 1, Paranebalia sp., and Ostracoda sp. 2 species have a higher resistance to oil contamination. Sediment reworking activity was also affected by oil addition. At the highest level of contamination, nearly no activity was observed due to the high mortality of macroorganisms. The results strongly suggest that an oil spill in this protected marine area with no previous history of contamination would have a deep impact on the non-adapted macrobenthic community

Fate of petroleum hydrocarbons in bioturbated pristine sediments from Caleta Valdés (Patagonia Argentina): An ex situ bioassay

Petroleum can pollute pristine shorelines as a consequence of accidental spills or chronic leaks. In this study, the fate of petroleum hydrocarbons in soft pristine sediment of Caleta Valdés (Argentina) subject toex situsimulated oil pollution was assessed. Sedimentary columns were exposed to medium and high concentrations of Escalante Crude Oil (ECO) and incubated in the laboratory during 30 days. Levels of aliphatic hydrocarbons at different depths of the sedimentary column were determined by gas chromatography. Oil penetration was limited to the first three centimetres in both treatments, and under this depth, hydrocarbons were clearly biogenic (terrestrial plants) as in the whole sedimentary column of the control assay. Bioturbation by macrobenthic infauna was strongly impacted by oil pollution which resulted in reduced sediment oxygenation and low burial of petroleum hydrocarbons. This may partly explain the limited hydrocarbon biodegradation observed, as indicated by the relatively high values of the ratios nC17/pristane,nC18/phytane, and total resolved aliphatic hydrocarbons/ unresolved complex mixture. Correspondingly, at the end of the experiment the most probable number of hydrocarbon-degrading bacteria reached ~ 10^3 MPN g−1 dry weight. These values were lower than those found in chronically polluted coastal sediments, reflecting a low activity level of the oil-degrading community. The results highlight the low attenuation capacities of Caleta Valdés pristine sediments to recover its original characteristics in a short time period if an oil spill occurs. In this work, we present a novel and integrative tool to evaluate the fate of petroleum hydrocarbons and their potential damage on pristine sediments

PhylArray: phylogenetic probe design algorithm for microarray

International audienceMOTIVATION: Microbial diversity is still largely unknown in most environments, such as soils. In order to get access to this microbial 'black-box', the development of powerful tools such as microarrays are necessary. However, the reliability of this approach relies on probe efficiency, in particular sensitivity, specificity and explorative power, in order to obtain an image of the microbial communities that is close to reality. RESULTS: We propose a new probe design algorithm that is able to select microarray probes targeting SSU rRNA at any phylogenetic level. This original approach, implemented in a program called 'PhylArray', designs a combination of degenerate and non-degenerate probes for each target taxon. Comparative experimental evaluations indicate that probes designed with PhylArray yield a higher sensitivity and specificity than those designed by conventional approaches. Applying the combined PhyArray/GoArrays strategy helps to optimize the hybridization performance of short probes. Finally, hybridizations with environmental targets have shown that the use of the PhylArray strategy can draw attention to even previously unknown bacteria

Towards the development of ecosystem-based indicators of mangroves functioning state in the context of the EU water framework directive

European Water Framework Directive is enforced in five tropical French Oversea Territories where mangroves are present. Developing bioindication tools to support the ecosystem-based management approach of the Directive is needed. A series of expert workshops was organized and led to the proposal of a strategy and of an applied research program to develop bioindication tools. The proceedings of the workshops are presented as a case study, as this is the first time such an integrative ecosystem-based approach is proposed in mangroves, combining structural and functional aspects, from forest structure to benthic community functioning

Impact of oil on bacterial community structure in bioturbated sediments

Oil spills threaten coastlines where biological processes supply essential ecosystem services. Therefore, it is crucial to understand how oil influences the microbial communities in sediments that play key roles in ecosystem functioning. Ecosystems such as sediments are characterized by intensive bioturbation due to burrowing macrofauna that may modify the microbial metabolisms. It is thus essential to consider the bioturbation when determining the impact of oil on microbial communities. In this study, an experimental laboratory device maintaining pristine collected mudflat sediments in microcosms closer to true environmental conditions - with tidal cycles and natural seawater - was used to simulate an oil spill under bioturbation conditions. Different conditions were applied to the microcosms including an addition of: standardized oil (Blend Arabian Light crude oil, 25.6 mg.g21 wet sediment), the common burrowing organism Hediste (Nereis) diversicolor and both the oil and H. diversicolor. The addition of H. diversicolor and its associated bioturbation did not affect the removal of petroleum hydrocarbons. After 270 days, 60% of hydrocarbons had been removed in all microcosms irrespective of the H. diversicolor addition. However, 16S-rRNA gene and 16S-cDNA T-RFLP and RT-PCR-amplicon libraries analysis showed an effect of the condition on the bacterial community structure, composition, and dynamics, supported by PerMANOVA analysis. The 16S-cDNA libraries from microcosms where H. diversicolor was added (oiled and un-oiled) showed a marked dominance of sequences related to Gammaproteobacteria. However, in the oiled-library sequences associated to Deltaproteobacteria and Bacteroidetes were also highly represented. The 16S-cDNA libraries from oiled-microcosms (with and without H. diversicolor addition) revealed two distinct microbial communities characterized by different phylotypes associated to known hydrocarbonoclastic bacteria and dominated by Gammaproteobacteria and Deltaproteobacteria. In the oiled-microcosms, the addition of H. diversicolor reduced the phylotype-richness, sequences associated to Actinobacteria, Firmicutes and Plantomycetes were not detected. These observations highlight the influence of the bioturbation on the bacterial community structure without affecting the biodegradation capacities

Caractérisation des communautés procaryotiques impliquées dans la bioremédiation d'un sol pollué par des hydrocarbures et développement d'outils d'analyse à haut débit

Environmental ecosystems are sensitive to damage from human activities and there is an increasing need to develop better methods for removing pollutants from soils. The removal of pollutants or their transformation to less toxic products by bioremediation is a less invasive and expensive process than classical decontamination. However, the use and optimization of bioremediation treatments requires knowledge on the microbial communities directly and indirectly involved in the degradation of the pollutants. Therefore, after the creation of a synthesis describing the microbial diversity and the available tools for the isolation and the identification of microorganisms, we have studied the structure and the dynamic of the bacterial communities during the bioremediation of an aliphatic hydrocarbon-polluted soil. Beyond the characterization of a bacterial "core-set" present throughout the decontamination process, we have identified several phylotypes potentially involved in degradation. In order to globally monitor these bacterial communities, we also have developed a probe design software (PhylArray) generating efficient and explorative probes for high-throughput molecular tools: the DNA microarrays.La bioremédiation est une réelle alternative aux techniques invasives de réhabilitation des sols pollués par des hydrocarbures. Cependant, la mise en place et l'optimisation de tels procédés nécessitent une meilleure connaissance des communautés microbiennes impliquées dans la biodégradation de ces polluants. Ainsi, nous avons étudié la structure et la dynamique des communautés bactériennes au cours d'un procédé de biostimulation d'un sol pollué aux hydrocarbures aliphatiques. Outre la caractérisation d' un core set bactérien présent tout au long du procédé, cette étude nous a permis d'identifier des phylotypes potentiellement impliqués dans la biodégradation de ces polluants. Enfin nous nous sommes attachés à mettre au point des outils moléculaires à haut débit, afin d'étudier plus globalement ces communautés environnementales, et ce par le développement d'un logiciel de sélection de sondes sensibles, spécifiques et exploratoires pour biopuces phylogénétiques (PhylArray

Caractérisation des communautés procaryotiques impliquées dans la bioremédiation d'un sol pollué par des hydrocarbures et développement d'outils d'analyse à haut débit (les biopuces ADN)

La bioremédiation est une réelle alternative aux techniques invasives de réhabilitation des sols pollués par des hydrocarbures. Cependant, la mise en place et l'optimisation de tels procédés nécessitent une meilleure connaissance des communautés microbiennes impliquées dans la biodégradation de ces polluants. Ainsi, nous avons étudié la structure et la dynamique des communautés bactériennes au cours d'un procédé de biostimulation d'un sol pollué aux hydrocarbures aliphatiques. Outre la caractérisation d' un core set bactérien présent tout au long du procédé, cette étude nous a permis d'identifier des phylotypes potentiellement impliqués dans la biodégradation de ces polluants. Enfin nous nous sommes attachés à mettre au point des outils moléculaires à haut débit, afin d'étudier plus globalement ces communautés environnementales, et ce par le développement d'un logiciel de sélection de sondes sensibles, spécifiques et exploratoires pour biopuces phylogénétiques (PhylArray)CLERMONT FD-BCIU Sci.et Tech. (630142101) / SudocSudocFranceF