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International audience; The fluctuation of environmental conditions drives the structure of microbial communities in estuaries, highly dynamic ecosystems. Microorganisms inhabiting estuarine sediments play a key role in ecosystem functioning. They are well adapted to the changing conditions, also threatened by the presence of pollutants. In order to determine the environmental characteristics driving the organization of the microbial assemblages, we conducted a seasonal survey along the Adour Estuary (Bay of Biscay, France) using 16S rRNA gene Illumina sequencing. Microbial diversity data were combined with a set of chemical analyses targeting metals and pharmaceuticals. Microbial communities were largely dominated by Proteobacteria (41 %) and Bacteroidota (32 %), showing a strong organization according to season, with an important shift in winter. The composition of microbial communities showed spatial distribution according to three main areas (upstream, middle, and downstream estuary) revealing the influence of the Adour River. Further analyses indicated that the microbial community was influenced by biogeochemical parameters (Corg/Norg and 13C) and micropollutants, including metals (As, Cu, Mn, Sn, Ti, and Zn) and pharmaceuticals (norfloxacin, oxolinic acid and trimethoprim). Network analysis revealed specific modules, organized around keystone taxa, linked to a pollutant type, providing information of paramount importance to understand the microbial ecology in estuarine ecosystems.

The land-sea continuum constitutes a mixing zone where soil microbial communities encounter, via runoff, those inhabiting marine coastal sediment resulting in community coalescence. Here, we propose an experimental approach, mimicking the land-sea continuum, to study the microbial community coalescence events in different situations, by 16S and 18S rRNA genes metabarcoding. The microbial community structure of sediment diverged with the soil inputs. For prokaryotes, phylogenetic enrichment and amplicon sequence variants (ASVs) replacements characterized the community changes in sediment receiving soil inputs. For fungi, despite phylogenetic enrichment was not observed, the fungal ASVs richness was maintained by soil inputs. Comparison of microbial communities revealed ASVs specific to sediment receiving soil inputs, and also ASVs shared with soil and/or runoff. Among these specific ASVs, four bacterial and one fungal ASVs were identified as indicators of coalescence. Our study provides evidences that coalescence involves the mixing of microorganisms and of the environment.

Dispersants, used for combating oil spills, increase hydrocarbon bioavailability promoting their biodegradation. Oil weathering process introduces harmful soluble hydrocarbons, such as polycyclic aromatic hydrocarbons (PAHs), into the water column, resulting in water-accommodated fraction (WAF). The presence of dispersants can influence the weathering process by increasing PAHs solubility, toxicity and biodegradability. However, little is known on how dispersants affect microbial communities and their degradation capacities, especially in cold environment where low temperature decreases microbial activity and thus hydrocarbon degradation. Here, we investigated the microbial community dynamics in cold water contaminated by WAF prepared from crude oil with or without a commercial dispersant (Finasol OSR52). The WAFs, prepared with Naphthenic North Atlantic crude oil, were used to contaminate seawater from Norwegian cold sites, one oil-contaminated and the other pristine. The WAF-contaminated seawaters were maintained in microcosms at 4 °C for 21 days. The content of PAHs and microbial compositions (16S rRNA gene sequencing) were determined at days 0, 7, 14 and 21. In addition, the 96 h toxicity assay with adult Acartia tonsa revealed WAFs toxicity at days 0 and 21. The toxicity of WAF mixtures, with and without dispersant, against Acartia tonsa was reduced during the experiment, but PAHs removal was not increased. The water from the oil-contaminated site showed the highest PAHs removal revealing legacy effect (presence of microorganisms adapted to PAHs). Additionally, our results reveal: i) microbial community plasticity allowing the adaptation to the presence of PAHs and dispersant, ii) specific bacteria taxa probably involved in PAHs degradation, and iii) dispersants shape the microbial communities dynamics by stimulating potential dispersant-degrading taxa, such as Fusibacter. Thus, our results provide valuable insights on the role of microbial community in determining the fate of water-solubilized hydrocarbon in cold environment while questioning the role of dispersant used for fighting oil spill.

International audience; This study aimed to determine the effect of the climatic change on the phototrophic communities of hypersaline microbial mats. Ocean acidification and warming were simulated alone and together on microbial mats placed into mesocosms. As expected, the temperature in the warming treatments increased by 4 °C from the initial temperature. Surprisingly, no significance difference was observed between the water pH of the different treatments despite of a decrease of 0.4 unit pH in the water reserves of acidification treatments. The salinity increased on the warming treatments and the dissolved oxygen concentration increased and was higher on the acidification treatments. A total of 37 pigments were identified belonging to chlorophylls, carotenes and xanthophylls families. The higher abundance of unknown chlorophyll molecules called chlorophyll derivatives was observed in the acidification alone treatment with a decrease in chlorophyll a abundance. This change in pigmentary composition was accompanied by a higher production of bound extracellular carbohydrates but didn't affect the photosynthetic efficiency of the microbial mats. A careful analysis of the absorption properties of these molecules indicated that these chlorophyll derivatives were likely bacteriochlorophyll c contained in the chlorosomes of green anoxygenic phototroph bacteria. Two hypotheses can be drawn from these results: 1/ the phototrophic communities of the microbial mats were modified under acidification treatment leading to a higher relative abundance of green anoxygenic bacteria, or 2/ the highest availability of CO2 in the environment has led to a shift in the metabolism of green anoxygenic bacteria being more competitive than other phototrophs.

The structure and diversity of microbial community inhabiting coastal sediments reflect the exposition to contaminants. Aiming to assess the changes in the microbiota from Sepetiba Bay (SB, Brazil) sediments, correlations between the 16S rRNA gene data (V4-V5 region), metal contamination factors (CF), and the ecological risk classification provided by the Quality Ratio (QR) index were considered. The results show that microbial diversity differs significantly between the less (SB external sector) and the most (SB internal sector) polluted sectors. Also, differences in the microbial community structure regarding the ecological risk classifications validated the QR index as a reliable tool to report the SB chronic contamination. Microbial indicator genera resistant to metals (Desulfatiglans, SEEP-SRB1, Spirochaeta 2, among others) presented mainly anaerobic metabolisms. These genera are related to the sulfate reducing and methanogenic metabolisms probably participating in the natural attenuation processes but also associated with greenhouse gas emissions. In contrast, microbial indicator genera sensitive to metals (Rubripirellula, Blastopirellula, Aquibacter, among others) presented mainly aerobic metabolisms. It is suggested that future works should investigate the metabolic functions to evaluate the influence of metallic contaminants on microbial community inhabiting SB sediment.

Large pilot scale bioremediation approaches were implemented for the treatments of oily sludge (OS) characterised by alkaline pH (pH > 9), high concentration of metals (3% dry weight) and high total petroleum hydrocarbons content (TPH) rangingbetween 22,000 and 67,300 mg kg −1 from a Tunisian petroleum refinery. The treatments included bioaugmentation and biostimulation approaches with autochthonous isolated bacterial strains and consortia. Chemical, microbial, and ecotoxicological analyses were performed over a period of 180 days incubation. The bioremediation treatments favoured the development of Proteobacteria, Firmicutes and Bacteroidetes following an ecological succession of specialist bacterial groups, first associated to hydrocarbon degradation (e.g. Marinobacter and Alcanivorax) that resulted in a greater extent of TPH-degradation (up to 80%), and the selection of metal resistant bacteria including Hyphomonas, Phaeobacter, and Desulfuromusa. The best performances were obtained when bioaugmentation and biostimulation were combined. Over 90% of the TPH initial concentration was degraded over 180 days, which was accompanied with a 3-fold reduction of ecotoxicity. Our study demonstrates the efficacy of large pilot scale bioremediation of highly contaminated oily sludge, providing the evidence that the management of autochthonous microbial communities is of paramount importance for the success of the bioremediation process.

Fungal microbiome interacts with the other biotic components in coastal sediment playing a key role in the overall coordination of the whole microbial community. These interactions are affected by human activities, such as the constant affluence of polycyclic aromatic hydrocarbons (PAHs). Although fungi and bacteria interactions have been found to play a key role in PAH bioremediation in soil, the effect of PAHs on fungal diversity and their specific interactions with bacteria in coastal sediments are yet to be investigated. The understanding of fungal bacterial interactions under PAH contamination is critical for further bioremediation regarding the important fungal diversity observed in coastal sediment. Here, we investigated the fungal bacterial co-occurrence in PAH-contaminated sediments. The co-occurrence network, constructed with sequencing data (bacterial 16S and fungal 18S rRNA genes barcoding) from 51 PAH-contaminated samples, revealed modules dominated by either fungi or bacteria, reflecting probably the different types of interaction possible between fungi and bacteria. Then, a network constructed from non-contaminated sample data was compared with a network built from the corresponding PAH-contaminated samples issued from a mesocosm experiment. The comparison revealed the effect of PAHs in fungi and bacteria interactions, characterized by a PAH-contaminated network exhibiting less abundant and diverse fungal and bacterial ASVs than the non-contaminated network. However, the links between the remaining ASVs in the PAH-contaminated network showed stronger correlations. Noteworthy, an ASV affiliated to Chrytridiomycota phylum was identified as a keystone fungal ASV forming a module in association with facultative anaerobic and anaerobic bacteria affiliated to the families Prolixibacteraceae, Fusobacteriaceae, and Desulfobulbaceae. These results suggest that fungi promote bacterial anaerobic metabolisms, which are important to cope with the presence of PAHs in sediments. Our study reveals the importance of fungal bacterial interactions in coastal sediments paving the way for future studies to fully understand fungal role in coastal sediment.

COnsensus DEgenerate Hybrid Oligonucleotide Primers (CODEHOP) were developed for the detection of the dsz B desulfinase gene (2′-hydroxybiphenyl-2-sulfinate desulfinase; EC 3.13.1.3) by polymerase chain reaction (PCR), which allow to reveal larger diversity than traditional primers. The new developed primers were used as molecular monitoring tool to drive a procedure for the isolation of desulfurizing microorganisms. The primers revealed a large dsz B gene diversity in environmental samples, particularly in diesel-contaminated soil that served as inoculum for enrichment cultures . The isolation procedure using the dibenzothiophene sulfone (DBTO 2) as sole sulfur source reduced drastically the dszB gene diversity. A dszB gene closely related to that carried by Gordonia species was selected. The desulfurization activity was confirmed by the production of desulfurized 2-hydroxybiphenyl (2-HBP). Metagenomic 16S rRNA gene sequencing showed that the Gordonia genus was represented at low abundance in the initial bacterial community. Such observation highlighted that the culture medium and conditions represent the bottleneck for isolating novel desulfurizing microorganisms. The new developed primers constitute useful tool for the development of appropriate cultural-dependent procedures, including medium and culture conditions, to access novel desulfurizing microorganisms useful for the petroleum industry.

Microbial communities from Sepetiba Bay (SB, Rio de Janeiro, Brazil), characterized by 16S rRNA gene (V4-V5 region) sequencing analysis, were found to be correlated with the metallic contamination factor and the Quality Ratio (QR) index. Consistently, the predicted function of microbial communities, obtained with Tax4Fun2, showed that the functional patterns in SB internal sector under the highest anthropogenic pressure were different from that observed in the external sector with the lowest contamination level. Signal transduction, cellular community, membrane transport, and energy metabolism were among the KEGG pathways favored by metallic contamination in the SB internal sector, while lipid metabolism, transcription, and translation were among the pathways favored in the SB external sector. Noteworthy, the relative proportions of KEGG pathways and genes associated with metallic homeostasis showed significant differences according to the SB sectors, consistently with the ecological risk classification (QR index) of sediments. The functional prediction approach is an economically viable alternative and presents an overview of the main pathways/genes favored in the SB microbiota exposed to long-term pollution. In contrast, the microgAMBI, ecological status index based on bacterial community composition, was not consistent with the metallic contamination of SB, suggesting that this index requires improvements to be applied in tropical areas. Our study also revealed a strong correlation between metal resistance genes (MRG) and antibiotic resistance genes (ARG), indicating that MRG and ARG are co-selected by the metallic contamination prevailing in SB.

International audience; In order to understand the effect of human practices on microbial mats organisation, the study aimed to investigate the biodiversity within microbial mats from exploited and abandoned salterns. Despite several attempts, archaeal 16S rRNA gene fragment sequences were not obtained, indicating that microbial mats were probably dominated by Bacteria with very low abundance of Archaea (< 1%). Thus, the study compared the bacterial and meiofaunal diversity of microbial mats from abandoned and exploited salterns. The higher salinity (101 ± 3.7 psu vs. 51.1 ± 0.7 psu; Welch t-test p < 0.05) of the exploited site maintained lower bacterial diversity in comparison to the abandoned site where the salinity gradient was no longer maintained. However, the microbial mats exhibited similar bacterial class composition while the eukaryotic diversity was significantly higher in the exploited saltern. The abandoned saltern was dominated by sulfate-reducing bacteria and Nematoda, while the exploited saltern was characterized by the presence of halophilic bacteria belonging to Marinobacter, Salinivibrio and Rhodohalobacter genera, and the larger abundance of Hypotrichia (ciliates). Such bacterial and eukaryotic diversity difference might be explained by human actions for salt recovery in exploited salterns such as scraping the surface of microbial mat and increasing salinity renewing the microbial mat each year. Such action decreases the bacterial diversity changing the food web structure that favour the presence of a larger diversity of eukaryotic organisms. Our study provides new insights on microbial mat communities inhabiting salterns, especially the consequences of abandoning saltern exploitation.

International audience; The cultivable fungal diversity from PAH-contaminated sediments was examined for the tolerance to polycyclic aromatic hydrocarbon (PAH). The 85 fungal strains, isolated in non-selective media, revealed a large diversity by ribosomal internal transcribed spacer (ITS) sequencing, even including possible new species. Most strains (64%) exhibited PAH-tolerance, indicating that sediments retain diverse cultivable PAH-tolerant fungi. The PAH-tolerance was linked neither to a specific taxon nor to the peroxidase genes (LiP, MnP and Lac). Examining the PAH-removal (degradation and/or sorption), Alternaria destruens F10.81 showed the best capacity with above 80% removal for phenanthrene, pyrene and fluoranthene, and around 65% for benzo[a]pyrene. A. destruens F10.81 internalized pyrene homogenously into the hyphae that contrasted with Fusarium pseudoygamai F5.76 in which PAH-vacuoles were observed but PAH removal was below 20%. Thus, our study paves the way for the exploitation of fungi in remediation strategies to mitigate the effect of PAH in coastal marine sediments.

Microbial communities inhabiting soil-water-sediment continuum in coastal areas provide important ecosystem services. Their adaptation in response to environmental stressors, particularly mitigating the impact of pollutants discharged from human activities, has been considered for the development of microbial biomonitoring tools, but their use is still in the infancy. Here, chemical and molecular (16S rRNA gene metabarcoding) approaches were combined in order to determine the impact of pollutants on microbial assemblages inhabiting the aquatic network of a soil-water-sediment continuum around the Ichkeul Lake (Tunisia), an area highly impacted by human activities. Samples were collected within the soil-river-lake continuum at three stations in dry (summer) and wet (winter) seasons. The contaminant pressure index (PI), which integrates Polycyclic aromatic hydrocarbons (PAHs), alkanes, Organochlorine pesticides (OCPs) and metal contents, and the microbial pressure index microgAMBI, based on bacterial community structure, showed significant correlation with contamination level and differences between seasons. The comparison of prokaryotic communities further revealed specific assemblages for soil, river and lake sediments. Correlation analyses identified potential "specialist" genera for the different compartments, whose abundances were correlated with the pollutant type found. Additionally, PICRUSt analysis revealed the metabolic potential for pollutant transformation or degradation of the identified "specialist" species, providing information to estimate the recovery capacity of the ecosystem. Such findings offer the possibility to define a relevant set of microbial indicators for assessing the effects of human activities on aquatic ecosystems. Microbial indicators, including the detection of “specialist” and sensitive taxa, and their functional capacity, might be useful, in combination with integrative microbial indices, to constitute accurate biomonitoring tools for the management and restoration of complex coastal aquatic systems.

International audience; This work aimed to evaluate the effect of acid mist on microbial communities developed on mortar with same composition from the wall of the National Museum in Rio de Janeiro. Five autochthonous microbial groups were inoculated on coated with acrylic paint and uncoated mortar surfaces and submitted to acid mist or water mist for 100 days. A heterogeneous biofilm was well developed in uncoated coupons under acid mist. Molecular analysis revealed that the bacterial communities were structured according to the incubation condition. The impact of coating on bacterial community structure was more important than the acid mist effect. The eukaryotic community structure was not affected by the acid mist, whereas the impact of coating by paint has been observed. The alpha diversity indexes also varied according to the treatment. H’ and D were higher for uncoated conditions, particularly when submitted to acid mist. Actinobacteria, Chloroflexi, Cyanobacteria, Firmicutes and Proteobacteria were dominant. Additionally, eukaryotic community was not affected by the acid mist and paint and Tremellomycetes dominated.

International audience; We described the bacterial diversity of walnut grove soils under organic and conventional farming. The bacterial communities of rhizospheric and non‐rhizospheric soils of pecan tree (Carya illinoensis K. Koch) were compared considering two phenological stages (sprouting and ripening). Sixteen OTUs were identified significantly more abundant according to the plant development, only one according to the farming condition, and none according to the soil origin. The OTUs specificaly abundant according to plant development included Actinobateria (2) and Betaproteobacteria (1) related OTUs more abundant at the sprouting stage, while at the fruit ripening stage the more abundant OTUs were related to Actinobacteria (6), Alphaproteobacteria (6), and unclassified Bacteria (1). The Gaiellaceae OTU18 (Actinobacteria) was more abundant under conventional farming. Thus, our study revealed that the plant development stage was the main factor shaping the bacterial community structure, while less influence was noticed for the farming condition. The bacterial communities exhibited specific metabolic capacities, a large range of carbon sources being used at the fruit ripening stage. The identified OTUs specifically more abundant represent indicators providing useful information on soil condition, potential tools for the management of soil bacterial communities.

International audience; Bacterial mono- and dialkyl glycerol ether lipids (MAGEs and DAGEs) are potential powerful lipid biomarkers whose respective diagenetic fate still needs to be precisely assessed. To do so, the lipid content of cultures of an anaerobic bacterium synthesizing both MAGEs and DAGEs was analyzed at different growth stages and after long-term incubation under aerobic or anaerobic conditions. The proportions of the different lipid classes (fatty acids, MAGEs and DAGEs), as well as their structural distributions, did not significantly vary during anaerobic growth under optimal conditions until the stationary phase. Long-term exposure to oxygen (up to 480 days) induced the preferential degradation of fatty acids and MAGEs relative to DAGEs and of sn-1 relative to sn-2 positional isomers of MAGEs. The degradation of fatty acids and MAGEs appeared slower under anaerobic conditions and no specific structures appeared preferentially degraded. All DAGE structures were very well preserved whatever the redox conditions and the time of incubation considered (480 days and 13 years under oxic and anoxic conditions, respectively). The results highlight the excellent diagenetic stability of DAGEs and the much lower potential of preservation of MAGEs, warranting caution for an eventual use of the latter compounds as tracers of environmental or microbial community changes.

2017-410 Mangroves are among the most sensitive marine ecosystems to oil pollution due both to the sensitivity of mangroves species and to the high persistence of hydrocarbons in these environments. Despite their ecological and socio-economic value, the potential effects of an oil spill on French Guiana mangroves remain so far unknown. Yet, there is an increasing transboundary risk of oil spill due to Brazilian offshore oil exploitation (in mid-April 2013, there were 122 offshore drilling rigs in Brazil, including 29 under construction – Brazilian Amapá region located in the French Guiana border area is thought to become an important world oil production area in the coming years). The aim of the PRISME project was to assess the natural degradation of oil in mangrove sediment as well as its impact on benthic communities (micro, meio and macrobenthos): a one-month in situ experiment was conducted in the young French Guianese mangrove (around 3 years old) at the mouth of the Sinnamary estuary. The experimental units consisted in eight plastic cores (Ø : 10 cm ; height: 30 cm) manually introduced within sediments. A thin layer of oiled sediment (2 cm, 20 000 ppm) was applied on four cores while the remaining four cores were considered as control (no oil addition). Three cores were additionally sampled at the beginning of the experiment as initial control sediments. After one month in situ, the eight cores were collected and sliced on site into different sedimentary layers aliquots for later analyses (hydrocarbons, bacterial, meio, macrofauna fauna diversity, bioturbation, biogeochemical parameters). Samples were sent to the different laboratories involved in this multidisciplinary project. Results and knowledge gained from this experimental work were used to develop an approach for assessing coastal vulnerability for oil spills preparedness in mangroves.

International audience; Bacterial communities inhabiting sediments in coastal areas endure the effect of strong anthropogenic pressure characterized by the presence of multiple contaminants. Understanding the effect of pollutants on the organization of bacterial communities is of paramount importance in order to unravel bacterial assemblages colonizing specific ecological niches. Here, chemical and molecular approaches were combined to investigate the bacterial communities inhabiting the sediments of the Ichkeul Lake/Bizerte Lagoon, a hydrological system under anthropogenic pressure. Although the microbial community of the Ichkeul Lake sediment was different to that of the Bizerte Lagoon, common bacterial genera were identified suggesting a lake-lagoon continuum probably due to the hydrology of the system exchanging waters according to the season. These genera represent bacterial “generalists” maintaining probably general biogeochemical functions. Linear discriminant analysis effect size (LEfSe) showed significant differential abundance distribution of bacterial genera according to the habitat, the pollution type and level. Further, correlation analyses identified specific bacterial genera which abundance was linked with pesticides concentrations in the lake, while in the lagoon the abundance of specific bacterial genera was found linked with the concentrations of PAHs (Polycyclic aromatic hydrocarbons) and organic forms of Sn. As well, bacterial genera which abundance was not correlated with the concentrations of pollutants were identified in both lake and lagoon. These findings represent valuable information, pointing out specific bacterial genera associated with pollutants, which represent assets for developing bacterial tools for the implementation, the management, and monitoring of bioremediation processes to mitigate the effect of pollutants in aquatic ecosystems.

International audience

International audience; Air samples from four contrasting outdoor environments including a park, an arable farm, a waste water treatment plant and a composting facility were analysed during the summer and winter months. The aim of the research was to study the feasibility of differentiating microbial communities from urban, rural and industrial areas between seasons with chemical and molecular markers such as microbial volatile organic compounds (MVOCs) and phospholipid fatty acids (PLFAs). Air samples (3 l) were collected every 2 h for a total of 6 h in order to assess the temporal variations of MVOCs and PLFAs along the day. MVOCs and VOCs concentrations varied over the day, especially in the composting facility which was the site where more human activities were carried out. At this site, total VOC concentration varied between 80 and 170 μg m−3 in summer and 20–250 μg m−3 in winter. The composition of MVOCs varied between sites due to the different biological substrates including crops, waste water, green waste or grass. MVOCs composition also differed between seasons as in summer they are more likely to get modified by oxidation processes in the atmosphere and in winter by reduction processes. The composition of microbial communities identified by the analysis of PLFAs also varied among the different locations and between seasons. The location with higher concentrations of PLFAs in summer was the farm (7297 ng m−3) and in winter the park (11,724 ng m−3). A specific set of MVOCs and PLFAs that most represent each one of the locations was identified by principal component analyses (PCA) and canonical analyses. Further to this, concentrations of both total VOCs and PLFAs were at least three times higher in winter than in summer. The difference in concentrations between summer and winter suggest that seasonal variations should be considered when assessing the risk of exposure to these compounds.

International audience; The environmental pollutions generated by human activities are important concerns that environmental risk assessment procedures have the purpose to evaluate and mitigate the effects. Microorganisms are among the first impacted by human generated pollutions. Furthermore, because they are essential actors in ecosystem functioning the evaluation of the pollution effects on microorganisms is of paramount importance. Their response may serve as proxy to report the effects on, and the recovering capacities of, the ecosystem. The behaviour of microorganisms in response to chemical pollution has been largely studied. In this chapter, we introduce the mechanisms underlying the microbial adaptation capacities involved in response to pollutants. We also discuss the basic knowledge inspiring microbial ecotoxicological tools reporting the pollutant effects that have been developed at the different biological organization levels, from genes and cellular processes to population and microbial community responses.

cited By 1; International audience; The objectives of this study were (1) to assess the responses of benthic nematodes to a polycyclic aromatic hydrocarbon (PAH) contamination and (2) to test bioremediation techniques for their efficiency in PAH degradation and their effects on nematodes. Sediments with their natural nematofauna communities from Bizerte lagoon (Tunisia) were subjected to a PAH mixture (100 ppm) of phenanthrene, fluoranthene, and pyrene during 30 days. Nematode abundance and diversity significantly decreased, and the taxonomic structure was altered. Results from multivariate analyses of the species abundance data revealed that PAH treatments were significantly different from the control. Spirinia parasitifera became the dominant species (70 % relative abundance) and appeared to be an “opportunistic” species to PAH contamination while Oncholaimus campylocercoides and Neochromadora peocilosoma were strongly inhibited. Biostimulation (addition of mineral salt medium) and bioaugmentation (inoculation of a hydrocarbonoclastic bacterium) were used as bioremediation techniques. Bioremediation treatments enhanced degradation of all three PAHs, with up to 96 % degradation for phenanthrene resulting in a significant stimulation of nematode abundance relative to control microcosms. Nevertheless, these treatments, especially the biostimulation provoked a weak impact on the community structure and diversity index relative to the control microcosms suggesting their feasibility in biorestoration of contaminated sediments.

Pitting corrosion in microalloyed steel and its welded joints occur in fabricated structures, such as pipelines for oil and gas transport, which are exposed to corrosive environments. Although the formation of the pits consumes only a relatively small mass of material, the role of the pits as defect sites for crack initiation and continuing corrosion can be substantial. Furthermore, in natural and industrial environments where microorganisms are widely distributed, pitting corrosion is aggravated by the release of many reactive microbial metabolites. This is a report of a 28-day study aimed at the microbiologically influenced corrosion of joint welds by gas tungsten arc welding (GTAW) and shielded metal arc welding (SMAW) exposed to laminar and turbulent flows. The characterization of the planktonic microbial community indicated that laminar flow created an anoxic environment promoting the dominance of anaerobic microorganisms, in particular sulfate reducing bacteria (SRB). Both types of welded joints showed similar pit densities in the base metal (BM), heat-affected zone (HAZ) and weld metal (WM), nevertheless the pit depth was higher in the HAZ than the BM and WM. On the other hand, turbulent flow retained dissolved oxygen content, ensuring the presence of aerobic microorganisms, but there was no inhibition of SRB growth. Although the pit depth was low in both types of welded joints, the pit density was exacerbated in only one HAZ of each welded joint due to WM reinforcement height, while the other HAZ showed low pit density. This study demonstrates that turbulent flow can reduce the biocorrosion of non-welded areas but can aggravate welded joints biocorrosion.

International audience; Worldwide, the corrosion of petroleum pipelines is the cause of severe operational failures and damages to the environment, and the majority of them may have been intensified by microbial activities. However, few scientific publications have investigated the influence of seawater with indigenous microorganisms in internal corrosion of welded joints in long-distance pipelines, which has motivated this study. Here, a combination of culture-based and molecular microbiological methods was applied to measure the microbial abundance, diversity and composition of biofilms formed on two corroded weldments (gas tungsten arc - GTAW and shielded metal arc - SMAW) of an API 5L X65 microalloyed steel under laminar and turbulent flow regimes using seawater from Guanabara Bay (Rio de Janeiro, Brazil). Overall, turbulent flow did not reduce the biomass of sessile microorganisms on welded joints compared to laminar flow. Phylogenetic analysis revealed that the Proteobacteria, Firmicutes and Bacteroidetes were the dominant phyla in biofilms attached to GTAW and SMAW joints. At the genus level, all samples showed similar microbial composition with dominance of Desulfovibrio. However, analysis at OTU level revealed that there were specific microorganisms in each corrosive sample, even though they were affiliated to sulfate-reducers. Thus, microbial community compositions were influenced by type of weld, flow regime and biofilm age. The results of our study will be of benefit to the further studies of weld biocorrosion and biofilm ecology within pipelines.

Summary Microbial diversity is generally considered as having no effect on the major processes of the ecosystem such as respiration or nutrient assimilation. However, information about the impact of diversity on minor functions such as xenobiotic degradation is scant. We studied the role of diversity on the capacity of an activated-sludge microbial community to eliminate phenanthrene, a polycyclic aromatic hydrocarbon. We also assessed the impact of diversity erosion on the ability of activated sludge to oxidize a wide range of organic compounds. The diversity of activated sludge was artificially modified by dilution to extinction followed by regrowth stage which led to communities with similar biomass but displaying a diversity gradient. The capacity of activated-sludge community to degrade phenanthrene was greatly modified: at high levels of diversity, the community was able to mineralize phenanthrene whereas at medium levels it first of all partially lost its ability to mineralize this pollutant and at the lowest diversity, the activated sludge completely lost its capacity to transform phenanthrene. Diversity depletion also reduced the metabolic diversity and biomass productivity of sewage-activated sludge. This study demonstrates that diversity erosion can greatly affect major ecosystem services such as pollutant removal.

In coastal marine sediment, oxygen fluctuations induced by bioturbating activities are widespread and exert a great influence, not only on the structure and diversity of the microbenthic communities, but also on their activities. Thus, the activity of benthic organisms can have a significant influence on the degradation of hydrocarbons (HC) and can favor the development of hydrocarbonoclastic microorganisms in contaminated marine sediments. Here, we have generated metatranscriptomic data from coastal marine sediments affected by oil addition and/or by the reworking activity of the marine polychaete Hediste diversicolor to gain insights into the active microbial groups involved in the response to oil addition under the oxygen-fluctuating conditions. The preliminary results suggest that the macrofauna promote the diversity of active aerobic hydrocarbonoclastic bacteria in marine sediments, even if its influence cannot be strongly observed at the microbial community expression profiles level.

Polycyclic aromatic hydrocarbons (PAHs) are widespread in marine ecosystems and originate from natural sources and anthropogenic activities. PAHs enter the marine environment in two main ways, corresponding to chronic pollution or acute pollution by oil spills. The global PAH fluxes in marine environments are controlled by the microbial degradation and the biological pump, which plays a role in particle settling and in sequestration through bioaccumulation. Due to their low water solubility and hydrophobic nature, PAHs tightly adhere to sediments leading to accumulation in coastal and deep sediments. Microbial assemblages play an important role in determining the fate of PAHs in water and sediments, supporting the functioning of biogeochemical cycles and the microbial loop. This review summarises the knowledge recently acquired in terms of both chronic and acute PAH pollution. The importance of the microbial ecology in PAH-polluted marine ecosystems is highlighted as well as the importance of gaining further in-depth knowledge of the environmental services provided by microorganisms., This review highlights the sources and fate of polycyclic aromatic hydrocarbons (PAHs) in the marine environment with particular emphasis on the microbial ecology and the biological pump controlling global PAH fluxes.

International audience; Bacterial non-isoprenoid alkyl glycerol ether lipids (AGEs) are widespread in the environment but little is known about their biological precursors and mode of formation as a function of varying growth condi- tions. Here, we searched for the presence of AGEs in five pure strains of mesophilic (growth temperature between 20 and 40 C) and heterotrophic sulfate reducing bacteria from the family Desulfobacteraceae grown on a wide range of hydrocarbons and organic acids as sole carbon and energy source. Gas chro- matography–mass spectrometry (GC–MS) analysis of hydrolyzed cells revealed the presence of mono- alkyl glycerols (MAGEs) in one Desulfatiferula species and of MAGEs and dialkyl glycerols (DAGEs) in two Desulfatibacillum species. Species from the genus Desulfococcus did not produce AGEs. The ether lipid composition appeared strongly dependent on the nature and the chain length of the carbon substrate, with few to > 50 homologues of AGEs formed from a single substrate. Growth on long chain n-alkyl com- pounds (C14 to C18 alk-1-enes and fatty acids) generated a much lower diversity of MAGEs and DAGEs than short chain substrates (nonanoate, octanoate and pyruvate) or isoprenoid alkenes (phytadienes). Such variation in alkyl glycerol production could be linked to the distinct pathways involved in the meta- bolism of the different substrates. Despite this substrate-dependent AGE composition, the average chain length and level of branching of each class of ether lipid remained remarkably stable whatever the growth substrate and the number of MAGE and DAGE homologues formed, indicating a compositional control of ether lipids by heterotrophic bacteria to maintain optimal membrane properties.

This study evaluates the in vitro impact of three bioremediation techniques: biostimulation, bioaugmentation, and a combination of biostimulation and bioaugmentation on microflora and nematofauna after an artificial contamination of la- goon sediments by anthracene. This sediment contamination tends to reduce the average load in aerobic heterotrophic bacte- ria in the water column, at the sediment interface and at the same time induces a highly significant decrease in the average densities of nematodes in all microcosms contaminated by this pollutant. After inoculation with the Staphylococcus sp. bac- terial strain, the increase in bacterial load was significant (p < 0.05) in the three compartments of anthracene-contaminated microcosms with relation to the controls. The addition of nitrates and phosphates to the contaminated microcosms generated a highly significant increase in the average concentration of free-living nematodes with relation to the controls.

A novel anaerobic, long-chain alkene-degrading, sulfate-reducing bacterium, strain LM2801T, was isolated from brackish sediment of a wastewater decantation facility of an oil refinery (Berre lagoon, France). Cells of strain LM2801T were Gram-negative, motile, slightly curved or vibrioid rods. Its optimum growth conditions were 30–36 °C, 6–10 g NaCl l−1 and pH 7.5. Strain LM2801T incompletely oxidized long-chain alkenes (from C14 to C23) and fatty acids (C14 to C24). The DNA G+C content was 45.5 mol%. Sequence analyses of the 16S rRNA and dsrAB genes indicated that the strain was a member of the family Desulfobacteraceae within the Deltaproteobacteria. This novel isolate possesses phenotypic and phylogenetic traits that do not allow its classification as a member of any previously described genus. Therefore, strain LM2801T is described as a member of a new genus, Desulfatiferula gen. nov., of which Desulfatiferula olefinivorans sp. nov. is the type species. The type strain of Desulfatiferula olefinivorans is LM2801T (=DSM 18843T =JCM 14469T).

International audience; 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.

Coastal shorelines suffer a variety of pollution injuries from both sides, from the sea, predominantly by crude oil from shipping and offshore mining, and from the land, principally through agricultural practices, urban wastewaters, and industrial activities. As a result, multi-pollution hot spots are found along shorelines, especially in estuaries and harbors. Microbial communities have evolved to adapt their metabolism to the presence of multi-contaminants (Duran et al. 2008; Gillan et al. 2005; Iannelli et al. 2012; Kaci et al. 2014; SabadiniSantos et al. 2014; Wang and Tam 2012). However, hydrocarbon compounds and crude oil-derived products are the most abundant pollutants, the more spectacular source being oil spill as illustrated by the recent catastrophe of Deepwater Horizon in 2010, the largest oil spill so far observed (Atlas and Hazen 2011). Although they are frequently found at unacceptably high concentrations, hydrocarbons are natural compounds, and thus, most of them can be biodegraded by the collective catabolic diversity of microorganisms (Duran and Goni Urriza 2010; Head et al. 2006; Leahy and Colwell 1990; Miralles et al. 2007; Paisse et al. 2011; Paisse et al. 2010), particularly demonstrated in coastal marine ecosystems such as salt marshes with microbial mat structures (Bordenave et al. 2004a; Bordenave et al. 2008; Bordenave et al. 2007; Bordenave et al. 2004b), mangroves (Brito et al. 2009; Brito et al. 2006) and estuaries (Chronopoulou et al. 2013; Coulon et al. 2012). However, coastal marine sediments constitute particular ecosystems submitted to fluctuating oxygenation and redox conditions from the tidal cycles and burrowing activities of the macrofauna that in turn drive microbial degradation processes (Cravo-Laureau and Duran 2014). The biodegradation of hydrocarbon compounds in such oxic/anoxic oscillating environments still remains poorly understood (Cravo-Laureau et al. 2011; Cuny et al. 2011; Vitte et al. 2013; Vitte et al. 2011). The understanding of the particular microbes involved, their ecology, their genetic and enzymatic capacities, their interactions, as well as their functioning in the changing redox conditions, is crucial for the implementation of efficient bioremediation strategies (Goni-Urriza et al. 2013; McGenity 2014). The DECAPAGE project (ANR CESA-2011-006 01; http://ipremeem.univ-pau.fr/live/DECAPAGE), funded by the French National Agency for Research (ANR), was precisely devoted to understand the adaptation mechanisms driving the reorganization of bacterial communities in response to petroleum in coastal sediments. In this special issue, the scientists involved in the DECAPAGE project present their contribution for understanding the microbial ecology in hydrocarbon-polluted mudflat sediments. The special issue also includes articles from scientists involved in other projects and programs in order to enlarge the topic to diverse ecosystems, to different biological organization levels from microbial and macrobenthic communities to bacterial populations, and addressing remediation strategies as well. Thus, the 15 articles compiled in this special issue explore diverse facets of the microbial ecology of hydrocarbon-polluted coastal sediments. The article of Acosta-Gonzalez et al. (2015) reviews the impact of the Prestige oil spill, an accident that occurred in 2002 at the Responsible editor: Philippe Garrigues

Bacterial glycerol ether lipids (alkylglycerols) have received increasing attention during the last decades, notably due to their potential role in cell resistance or adaptation to adverse environmental conditions. Major uncertainties remain, however, regarding the origin, biosynthesis, and modes of formation of these uncommon bacterial lipids. We report here the preponderance of monoalkyl- and dialkylglycerols (1-O-alkyl-, 2-O-alkyl-, and 1,2-O-dialkylglycerols) among the hydrolyzed lipids of the marine mesophilic sulfate-reducing proteobacterium Desulfatibacillum alkenivorans PF2803T grown on n-alkenes (pentadec-1-ene or hexadec-1-ene) as the sole carbon and energy source. Alkylglycerols account for one-third to two-thirds of the total cellular lipids (alkylglycerols plus acylglycerols), depending on the growth substrate, with dialkylglycerols contributing to one-fifth to two-fifths of the total ether lipids. The carbon chain distribution of the lipids of D. alkenivorans also depends on that of the substrate, but the chain length and methyl-branching patterns of fatty acids and monoalkyl- and dialkylglycerols are systematically congruent, supporting the idea of a biosynthetic link between the three classes of compounds. Vinyl ethers (1-alken-1'-yl-glycerols, known as plasmalogens) are not detected among the lipids of strain PF2803T. Cultures grown on different (per)deuterated n-alkene, n-alkanol, and n-fatty acid substrates further demonstrate that saturated alkylglycerols are not formed via the reduction of hypothetic alken-1'-yl intermediates. Our results support an unprecedented biosynthetic pathway to monoalkyl/monoacyl- and dialkylglycerols in anaerobic bacteria and suggest that n-alkyl compounds present in the environment can serve as the substrates for supplying the building blocks of ether phospholipids of heterotrophic bacteria.

International audience; The present study aimed to examine whether the use of dispersant would be suitable for favoring the hydrocarbon degradation in coastal marine sediments without impacting negatively micro- and macrobenthic organisms. Mudflat sediments, maintained during 286 days in mesocosms designed to simulate natural conditions, were contaminated or not with Ural blend crude oil (REBCO) and treated or not with third-generation dispersant (Finasol OSR52). While the dispersant did not lead to an increase of hydrocarbon biodegradation, its use enables an attenuation of more than 55 % of the sediment concentration of total petroleum hydrocarbons (TPH). Canonical correspondence analysis (CCA) correlating T-RFLP patterns with the hydrocarbon content and bacterial abundance indicated weak differences between the different treatments except for the mesocosm treated with oil and dispersant for which a higher bacterial biomass was observed. The use of the dispersant did not significantly decrease the macrobenthic species richness or macroorganisms' densities in uncontaminated or contaminated conditions. However, even if the structure of the macrobenthic communities was not affected, when used in combination with oil, biological sediment reworking coefficient was negatively impacted. Although the use of the dispersant may be worth considering in order to accelerate the attenuation of hydrocarbon-contaminated mudflat sediments, long-term effects on functional aspects of the benthic system such as bioturbation and bacterial activity should be carefully studied before.

International audience; The present study aimed to examine whether the physical reworking of sediments by harrowing would be suitable for favouring the hydrocarbon degradation in coastal marine sediments. Mudflat sediments were maintained in mesocosms under conditions as closer as possible to those prevailing in natural environments with tidal cycles. Sediments were contaminated with Ural blend crude oil, and in half of them, harrowing treatment was applied in order to mimic physical reworking of surface sediments. Hydrocarbon distribution within the sediment and its removal was followed during 286 days. The harrowing treatment allowed hydrocarbon compounds to penetrate the first 6 cm of the sediments, and biodegradation indexes (such as n-C18/phytane) indicated that biodegradation started 90 days before that observed in untreated control mesocosms. However, the harrowing treatment had a severe impact on benthic organisms reducing drastically the macrofaunal abundance and diversity. In the harrowing-treated mesocosms, the bacterial abundance, determined by 16S rRNA gene Q-PCR, was slightly increased; and terminal restriction fragment length polymorphism (T-RFLP) analyses of 16S rRNA genes showed distinct and specific bacterial community structure. Co-occurrence network and canonical correspondence analyses (CCA) based on T-RFLP data indicated the main correlations between bacterial operational taxonomic units (OTUs) as well as the associations between OTUs and hydrocarbon compound contents further supported by clustered correlation (ClusCor) analysis. The analyses highlighted the OTUs constituting the network structural bases involved in hydrocarbon degradation. Negative correlations indicated the possible shifts in bacterial communities that occurred during the ecological succession.

International audience; Microbial communities inhabiting coastal marine sediments at intertidal zones are subjected to fluctuating environmental conditions according to tide level and reworking activities of macroorganisms. Such fluctuations are important drivers for the microbial communities’ assemblages and organization that in turn determine the microbial hydrocarbon-degradation processes. Notwithstanding the difficulties to access the entire ecosystem complexity, experimental ecology studies, involving more or less sophisticated experimental systems, have demonstrated to be suitable approaches. Several systems have been developed to tackle the role of microorganisms in hydrocarbon degradation in marine sediments in order to decipher the role of microbial communities in hydrocarbon degradation and understand the microbial behavior in response to an oil spill. We describe here a mesocosm system that maintains sediments under conditions as close as possible to those prevailing in the environment. We explain how to collect mudflat sediments conserving their structure and how to apply crude oil in order to simulate an oil spill. Subsampling procedures are also provided for following the fate of crude oil hydrocarbons, the macrofauna burrowing activity, as well as the behavior and dynamic of microbial communities.

cited By 1; International audience; The behaviour of sulphate-reducing microbial community was investigated at the oxic–anoxic interface (0–2 cm) of marine sediments when submitted to oil and enhanced bioturbation activities by the addition of Hediste diversicolor. Although total hydrocarbon removal was not improved by the addition of H. diversicolor, terminal restriction fragment length polymorphism (T-RFLP) analyses based on dsrAB (dissimilatory sulphite reductase) genes and transcripts showed different patterns according to the presence of H. diversicolor which favoured the abundance of dsrB genes during the early stages of incubation. Complementary DNA (cDNA) dsrAB libraries revealed that in presence of H. diversicolor, most dsrAB sequences belonged to hydrocarbonoclastic Desulfobacteraceae, suggesting that sulphate-reducing microorganisms (SRMs) may play an active role in hydrocarbon biodegradation in sediments where the reworking activity is enhanced. Furthermore, the presence of dsrAB sequences related to sequences found associated to environments with high dinitrogen fixation activity suggested potential N2 fixation by SRMs in bioturbated-polluted sediments.

A novel marine sulfate-reducing bacterium, strain CV2803T, which is able to oxidize aliphatic hydrocarbons, was isolated from a hydrocarbon-polluted marine sediment (Gulf of Fos, France). The cells were rod-shaped and slightly curved, measuring 0·6×2·2–5·5 μm. Strain CV2803Tstained Gram-negative and was non-motile and non-spore-forming. Optimum growth occurred in the presence of 24 g NaCl l−1, at pH 7·5 and at a temperature between 28 and 35 °C. Strain CV2803Toxidized alkanes (from C13to C18) and alkenes (from C7to C23). The DNA G+C content was 41·4 mol%. Comparative sequence analyses of the 16S rRNA gene and dissimilatory sulfite reductase (dsrAB) gene and those of other sulfate-reducing bacteria, together with its phenotypic properties, indicated that strain CV2803Twas a member of a distinct cluster that contained unnamed species. Therefore, strain CV2803T(=DSM 15576T=ATCC BAA-743T) is proposed as the type strain of a novel species in a new genus,Desulfatibacillum aliphaticivoransgen. nov., sp. nov.

cited By 4; International audience; Sediments from Bizerta lagoon were used in an experimental microcosm setup involving three scenarios for the bioremediation of anthracene-polluted sediments, namely bioaugmentation, biostimulation, and a combination of both bioaugmentation and biostimulation. In order to investigate the effect of the biotreatments on the benthic biosphere, 16S rRNA gene-based T-RFLP bacterial community structure and the abundance and diversity of the meiofauna were determined throughout the experiment period. Addition of fresh anthracene drastically reduced the benthic bacterial and meiofaunal abundances. The treatment combining biostimulation and bioaugmentation was most efficient in eliminating anthracene, resulting in a less toxic sedimentary environment, which restored meiofaunal abundance and diversity. Furthermore, canonical correspondence analysis showed that the biostimulation treatment promoted a bacterial community favorable to the development of nematodes while the treatment combining biostimulation and bioaugmentation resulted in a bacterial community that advantaged the development of the other meiofauna taxa (copepods, oligochaetes, polychaetes, and other) restoring thus the meiofaunal structure. The results highlight the importance to take into account the bacteria/meiofauna interactions during the implementation of bioremediation treatment.