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1.
Extremophiles ; 22(5): 825, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29492667

RESUMO

In the original publication there is a mistake in the supplementary material. The correct supplementary material is provided in this correction article.

2.
Extremophiles ; 22(1): 13-27, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29067531

RESUMO

Over the last decades, there has been growing interest about the ecological role of hydrothermal sulfide chimneys, their microbial diversity and associated biotechnological potential. Here, we performed dual-index Illumina sequencing of bacterial and archaeal communities on active and inactive sulfide chimneys collected from the Kolumbo hydrothermal field, situated on a geodynamic convergent setting. A total of 15,701 OTUs (operational taxonomic units) were assigned to 56 bacterial and 3 archaeal phyla, 133 bacterial and 16 archaeal classes. Active chimney communities were dominated by OTUs related to thermophilic members of Epsilonproteobacteria, Aquificae and Deltaproteobacteria. Inactive chimney communities were dominated by an OTU closely related to the archaeon Nitrosopumilus sp., and by members of Gammaproteobacteria, Deltaproteobacteria, Planctomycetes and Bacteroidetes. These lineages are closely related to phylotypes typically involved in iron, sulfur, nitrogen, hydrogen and methane cycling. Overall, the inactive sulfide chimneys presented highly diverse and uniform microbial communities, in contrast to the active chimney communities, which were dominated by chemolithoautotrophic and thermophilic lineages. This study represents one of the most comprehensive investigations of microbial diversity in submarine chimneys and elucidates how the dissipation of hydrothermal activity affects the structure of microbial consortia in these extreme ecological niches.


Assuntos
Fontes Hidrotermais/microbiologia , Microbiota , Sulfetos/análise , Archaea/classificação , Archaea/genética , Archaea/isolamento & purificação , Archaea/metabolismo , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Hidrogênio/metabolismo , Fontes Hidrotermais/química , Ferro/metabolismo , Metano/metabolismo , Nitrogênio/metabolismo , Sulfetos/metabolismo
3.
Anal Bioanal Chem ; 409(19): 4539-4549, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28540460

RESUMO

Chlorophyll a (Chl a) is the predominant pigment in every single photosynthesizing organism including phytoplankton and one of the most commonly measured water quality parameters. Various methods are available for Chl a analysis, but the majority of them are of limited throughput and require considerable effort and time from the operator. The present study describes a high-throughput, microplate-based fluorometric assay for rapid quantification of Chl a in phytoplankton extracts. Microplate sealing combined with ice cooling was proved an effective means for diminishing solvent evaporation during sample loading and minimized the analytical errors involved in Chl a measurements with a fluorescence microplate reader. A set of operating parameters (settling time, detector gain, sample volume) were also optimized to further improve the intensity and reproducibility of Chl a fluorescence signal. A quadratic regression model provided the best fit (r 2 = 0.9998) across the entire calibration range (0.05-240 pg µL-1). The method offered excellent intra- and interday precision (% RSD 2.2 to 11.2%) and accuracy (% relative error -3.8 to 13.8%), while it presented particularly low limits of detection (0.044 pg µL-1) and quantification (0.132 pg µL-1). The present assay was successfully applied on marine phytoplankton extracts, and the overall results were consistent (average % relative error -14.8%) with Chl a concentrations (including divinyl Chl a) measured by high-performance liquid chromatography (HPLC). More importantly, the microplate-based method allowed the analysis of 96 samples/standards within a few minutes, instead of hours or days, when using a traditional cuvette-based fluorometer or an HPLC system. Graphical abstract TChl a concentrations (i.e. sum of Chl a and divinyl Chl a in ng L-1) measured in seawater samples by HPLC and fluorescence microplate reader.


Assuntos
Clorofila/análise , Fitoplâncton/química , Clorofila A , Cromatografia Líquida de Alta Pressão , Limite de Detecção , Espectrometria de Fluorescência
4.
Environ Microbiol ; 18(4): 1122-36, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26487573

RESUMO

Hydrothermal vents represent a deep, hot, aphotic biosphere where chemosynthetic primary producers, fuelled by chemicals from Earth's subsurface, form the basis of life. In this study, we examined microbial mats from two distinct volcanic sites within the Hellenic Volcanic Arc (HVA). The HVA is geologically and ecologically unique, with reported emissions of CO2 -saturated fluids at temperatures up to 220°C and a notable absence of macrofauna. Metagenomic data reveals highly complex prokaryotic communities composed of chemolithoautotrophs, some methanotrophs, and to our surprise, heterotrophs capable of anaerobic degradation of aromatic hydrocarbons. Our data suggest that aromatic hydrocarbons may indeed be a significant source of carbon in these sites, and instigate additional research into the nature and origin of these compounds in the HVA. Novel physiology was assigned to several uncultured prokaryotic lineages; most notably, a SAR406 representative is attributed with a role in anaerobic hydrocarbon degradation. This dataset, the largest to date from submarine volcanic ecosystems, constitutes a significant resource of novel genes and pathways with potential biotechnological applications.


Assuntos
Archaea/classificação , Archaea/genética , Bactérias/classificação , Bactérias/genética , Ecossistema , Fontes Hidrotermais/microbiologia , Archaea/isolamento & purificação , Bactérias/isolamento & purificação , Sequência de Bases , Geologia , Metagenômica , RNA Ribossômico 16S/genética , Temperatura
5.
Environ Sci Technol ; 47(12): 6449-56, 2013 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-23668287

RESUMO

Two-dimensional compound-specific isotope analysis (2D-CSIA), combining stable carbon and chlorine isotopes, holds potential for monitoring of natural attenuation of chlorinated ethenes (CEs) in contaminated soil and groundwater. However, interpretation of 2D-CSIA data sets is challenged by a shortage of experimental Cl isotope enrichment factors. Here, isotope enrichments factors for C and Cl (i.e., εC and εCl) were determined for biodegradation of tetrachloroethene (PCE) and trichloroethene (TCE) using microbial enrichment cultures from a heavily CE-contaminated aquifer. The obtained values were εC = -5.6 ± 0.7‰ (95% CI) and εCl = -2.0 ± 0.5‰ for PCE degradation and εC = -8.8 ± 0.2‰ and εCl = -3.5 ± 0.5‰ for TCE degradation. Combining the values for both εC and εCl yielded mechanism-diagnostic εCl/εC ratios of 0.35 ± 0.11 and 0.37 ± 0.11 for the degradation of PCE and TCE, respectively. Application of the obtained εC and εCl values to a previously investigated field site gave similar estimates for the fraction of degraded contaminant as in the previous study, but with a reduced uncertainty in assessment of the natural attenuation. Furthermore, 16S rRNA gene clone library analyses were performed on three samples from the PCE degradation experiments. A species closely related to Desulfitobacterium aromaticivorans UKTL dominated the reductive dechlorination process. This study contributes to the development of 2D-CSIA as a tool for evaluating remediation strategies of CEs at contaminated sites.


Assuntos
Poluentes Atmosféricos/química , Biodegradação Ambiental , Isótopos de Carbono/química , Cloro/química , Monitoramento Ambiental/métodos , Isótopos/química , Tricloroetileno/química
6.
Front Microbiol ; 14: 1188544, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37455712

RESUMO

Introduction: Active hydrothermal vents of volcanic origin provide a remarkable manifestation of life on Earth under extreme conditions, which may have consequences for our understanding of habitability on other terrestrial bodies as well. Methods: Here, we performed for the first time Illumina sequencing of bacterial and archaeal communities on sub-seafloor samples collected from the Santorini-Kolumbo volcanic field. A total of 19 (3-m long) gravity corers were collected and processed for microbial community analysis. Results: From a total of 6,46,671 produced V4 sequences for all samples, a total of 10,496 different Operational Taxonomic Units (OTUs) were identified that were assigned to 40 bacterial and 9 archaeal phyla and 14 candidate divisions. On average, the most abundant phyla in all samples were Chloroflexi (Chloroflexota) (24.62%), followed by Proteobacteria (Pseudomonadota) (11.29%), Firmicutes (Bacillota) (10.73%), Crenarchaeota (Thermoproteota) (8.55%), and Acidobacteria (Acidobacteriota) (8.07%). At the genus level, a total of 286 known genera and candidate genera were mostly dominated by members of Bacillus, Thermoflexus, Desulfatiglans, Pseudoalteromonas, and Pseudomonas. Discussion: In most of the stations, the Chao1 values at the deeper layers were comparable to the surface sediment samples denoting the high diversity in the subsurface of these ecosystems. Heatmap analysis based on the 100 most abundant OTUs, grouped the sampling stations according to their geographical location, placing together the two hottest stations (up to 99°C). This result indicates that this specific area within the active Kolumbo crater create a distinct niche, where microorganisms with adaptation strategies to withstand heat stresses can thrive, such as the endospore-forming Firmicutes.

7.
Nature ; 440(7081): 203-7, 2006 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-16525471

RESUMO

The chemical composition of the Bannock basin has been studied in some detail. We recently showed that unusual microbial populations, including a new division of Archaea (MSBL1), inhabit the NaCl-rich hypersaline brine. High salinities tend to reduce biodiversity, but when brines come into contact with fresher water the natural haloclines formed frequently contain gradients of other chemicals, including permutations of electron donors and acceptors, that may enhance microbial diversity, activity and biogeochemical cycling. Here we report a 2.5-m-thick chemocline with a steep NaCl gradient at 3.3 km within the water column betweeen Bannock anoxic hypersaline brine and overlying sea water. The chemocline supports some of the most biomass-rich and active microbial communities in the deep sea, dominated by Bacteria rather than Archaea, and including four major new divisions of Bacteria. Significantly higher metabolic activities were measured in the chemocline than in the overlying sea water and underlying brine; functional analyses indicate that a range of biological processes is likely to occur in the chemocline. Many prokaryotic taxa, including the phylogenetically new groups, were confined to defined salinities, and collectively formed a diverse, sharply stratified, deep-sea ecosystem with sufficient biomass to potentially contribute to organic geological deposits.


Assuntos
Archaea/metabolismo , Bactérias/metabolismo , Ecossistema , Oxigênio/metabolismo , Células Procarióticas/metabolismo , Água do Mar/microbiologia , Microbiologia da Água , Aerobiose , Anaerobiose , Archaea/classificação , Archaea/genética , Archaea/isolamento & purificação , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Biodiversidade , Dados de Sequência Molecular , Oceanos e Mares , Células Procarióticas/classificação , Navios
8.
Proc Natl Acad Sci U S A ; 106(23): 9151-6, 2009 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-19470485

RESUMO

Urania basin in the deep Mediterranean Sea houses a lake that is >100 m deep, devoid of oxygen, 6 times more saline than seawater, and has very high levels of methane and particularly sulfide (up to 16 mM), making it among the most sulfidic water bodies on Earth. Along the depth profile there are 2 chemoclines, a steep one with the overlying oxic seawater, and another between anoxic brines of different density, where gradients of salinity, electron donors and acceptors occur. To identify and differentiate the microbes and processes contributing to the turnover of organic matter and sulfide along the water column, these chemoclines were sampled at a high resolution. Bacterial cell numbers increased up to a hundredfold in the chemoclines as a consequence of elevated nutrient availability, with higher numbers in the upper interface where redox gradient was steeper. Bacterial and archaeal communities, analyzed by DNA fingerprinting, 16S rRNA gene libraries, activity measurements, and cultivation, were highly stratified and metabolically more active along the chemoclines compared with seawater or the uniformly hypersaline brines. Detailed analysis of 16S rRNA gene sequences revealed that in both chemoclines delta- and epsilon-Proteobacteria, predominantly sulfate reducers and sulfur oxidizers, respectively, were the dominant bacteria. In the deepest layers of the basin MSBL1, putatively responsible for methanogenesis, dominated among archaea. The data suggest that the complex microbial community is adapted to the basin's extreme chemistry, and the elevated biomass is driven largely by sulfur cycling and methanogenesis.


Assuntos
Archaea/metabolismo , Bactérias/metabolismo , Água do Mar/microbiologia , Enxofre/metabolismo , Ecossistema , Manganês/metabolismo , Dados de Sequência Molecular , Nitratos/metabolismo , Oxigênio/metabolismo , Salinidade , Água/metabolismo
9.
Front Microbiol ; 13: 1060168, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36687571

RESUMO

Introduction: Shallow hydrothermal systems share many characteristics with their deep-sea counterparts, but their accessibility facilitates their study. One of the most studied shallow hydrothermal vent fields lies at Paleochori Bay off the coast of Milos in the Aegean Sea (Greece). It has been studied through extensive mapping and its physical and chemical processes have been characterized over the past decades. However, a thorough description of the microbial communities inhabiting the bay is still missing. Methods: We present the first in-depth characterization of the prokaryotic communities of Paleochori Bay by sampling eight different seafloor types that are distributed along the entire gradient of hydrothermal influence. We used deep sequencing of the 16S rRNA marker gene and complemented the analysis with qPCR quantification of the 16S rRNA gene and several functional genes to gain insights into the metabolic potential of the communities. Results: We found that the microbiome of the bay is strongly influenced by the hydrothermal venting, with a succession of various groups dominating the sediments from the coldest to the warmest zones. Prokaryotic diversity and abundance decrease with increasing temperature, and thermophilic archaea overtake the community. Discussion: Relevant geochemical cycles of the Bay are discussed. This study expands our limited understanding of subsurface microbial communities in acidic shallow-sea hydrothermal systems and the contribution of their microbial activity to biogeochemical cycling.

10.
Sci Rep ; 11(1): 1336, 2021 01 14.
Artigo em Inglês | MEDLINE | ID: mdl-33446715

RESUMO

Although the rise of antibiotic and multidrug resistant bacteria is one of the biggest current threats to human health, our understanding of the mechanisms involved in antibiotic resistance selection remains scarce. We performed whole genome sequencing of 21 Pseudomonas strains, previously isolated from an active submarine volcano of Greece, the Kolumbo volcano. Our goal was to identify the genetic basis of the enhanced co-tolerance to antibiotics and acidity of these Pseudomonas strains. Pangenome analysis identified 10,908 Gene Clusters (GCs). It revealed that the numbers of phage-related GCs and sigma factors, which both provide the mechanisms of adaptation to environmental stressors, were much higher in the high tolerant Pseudomonas strains compared to the rest ones. All identified GCs of these strains were associated with antimicrobial and multidrug resistance. The present study provides strong evidence that the CO2-rich seawater of the volcano associated with low pH might be a reservoir of microorganisms carrying multidrug efflux-mediated systems and pumps. We, therefore, suggest further studies of other extreme environments (or ecosystems) and their associated physicochemical parameters (or factors) in the rise of antibiotic resistance.


Assuntos
Adaptação Fisiológica/genética , Farmacorresistência Bacteriana Múltipla/genética , Genoma Bacteriano , Fontes Hidrotermais/microbiologia , Pseudomonas/genética , Água do Mar/microbiologia , Grécia , Pseudomonas/isolamento & purificação
11.
Int Microbiol ; 13(3): 143-50, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20890848

RESUMO

The variability of bacterial community composition and diversity was studied by comparative analysis of five 16S rRNA gene clone libraries from deep-sea sediments (water column depth: 4000 m) of the Northeastern Pacific Ocean and Eastern Mediterranean Sea. This is the first comparison of the bacterial communities living in these deep-sea ecosystems. The estimated chlorophyll a, organic carbon, and C/N ratio provided evidence of significant differences in the trophic state of the sediments between the Northeastern Pacific Ocean and the much warmer Eastern Mediterranean Sea. A diverse range of 16S rRNA gene phylotypes was found in the sediments of both regions. These were represented by 11 different taxonomic groups, with Gammaproteobacteria predominating in the Northeastern Pacific Ocean sediments and Acidobacteria in the Eastern Mediterranean microbial community. In addition, several 16S rRNA gene phylotypes only distantly related to any of the previously identified sequences (non-affiliated rRNA genes) represented a significant fraction of the total sequences. The potential diversity at the two sites differs but remains largely unexplored and remains of continuing scientific interest.


Assuntos
Bactérias/classificação , Bactérias/genética , Biodiversidade , Sedimentos Geológicos/microbiologia , Metagenoma , Bactérias/química , Carbono/análise , Clorofila/análise , Clorofila A , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , DNA Ribossômico/química , DNA Ribossômico/genética , Mar Mediterrâneo , Dados de Sequência Molecular , Compostos de Nitrogênio/análise , Oceano Pacífico , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Syst Appl Microbiol ; 32(1): 17-26, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19058941

RESUMO

This study is the first culture-independent report on the regional variability of bacterial diversity in oxic sediments from the unexplored southern Cretan margin (SCM). Three main deep basins (water column depths: 2670-3603m), located at the mouth of two submarine canyons (Samaria Gorge and Paximades Channel) and an adjacent slope system, as well as two shallow upper-slope stations (water column depths: 215 and 520m), were sampled. A total of 454 clones were sequenced and the bacterial richness, estimated through five clone libraries using rarefaction analysis, ranged from 71 to 296 unique phylotypes. The average sequence identity of the retrieved Cretan margin sequences compared to the >1,000,000 known rRNA sequences was only 93.5%. A diverse range of prokaryotes was found in the sediments, which were represented by 15 different taxonomic groups at the phylum level. The phylogenetic analysis revealed that these new sequences grouped with the phyla Acidobacteria, Planctomycetes, Actinobacteria, Gamma-, Alpha- and Delta-proteobacteria. Only a few bacterial clones were affiliated with Chloroflexi, Bacteroidetes, Firmicutes, Gemmatimonadetes, Verrucomicrobia, Nitrospirae, Beta-proteobacteria, Lentisphaerae and Dictyoglomi. A large fraction of the retrieved sequences (12%) did not fall into any taxonomic division previously characterized by molecular criteria, whereas four novel division-level lineages, termed candidate division SCMs, were identified. Bacterial community composition demonstrated significant differences in comparison to previous phylogenetic studies. This divergence was mainly triggered by the dominance of Acidobacteria and Actinobacteria and reflected a bacterial community different from that currently known for oxic and pristine marine sediments.


Assuntos
Bactérias/classificação , Variação Genética , Sedimentos Geológicos/microbiologia , Filogenia , Actinobacteria/classificação , Actinobacteria/genética , Bactérias/genética , Clonagem Molecular , Análise por Conglomerados , DNA Ribossômico , Ecossistema , Biblioteca Gênica , Grécia , Mar Mediterrâneo , Proteobactérias/classificação , Proteobactérias/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
13.
Mar Environ Res ; 146: 101-108, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30929836

RESUMO

Assessing the status of important carbon sinks such as seagrass meadows is of primary importance when dealing with potential climate change mitigation strategies. This study examined plant and sediment properties in seagrass meadows (Cymodocea nodosa (Ucria) Asch.) from two high pCO2-low pH Mediterranean vent systems, located at Milos (Greece) and Vulcano (Italy) Islands, providing insights on carbon storage potential in future acidified oceans. Contrary to what has been suggested, carbon content (both inorganic and organic) and its surficial accumulation decreased at high pCO2-low pH in comparison with controls. The decrease in inorganic carbon may result from the higher solubility of carbonates due to the more acidic conditions. At Vulcano, the seagrass properties (e.g., leaf area, biomass) appeared negatively affected by environmental conditions at high pCO2-low pH conditions and this may have had a detrimental effect on the organic carbon content and accumulation. At Milos, organic carbon decreased at high pCO2-low pH conditions, despite the increase in seagrass aboveground biomass, leaf length and area, probably as a consequence of site-specific features, which need further investigation and may include both biotic and abiotic factors (e.g., oligotrophic conditions, decreased sedimentation rate and input of allochthonous material). Results suggest that, in contrast to previous predictions based exclusively on the expected positive response of seagrasses to ocean acidification, carbon storage capacity of the seagrass C. nodosa may not increase at high pCO2-low pH conditions. This study emphasizes the need to investigate further the potential alteration in the climate mitigation service delivered by seagrass meadows in acidified oceans.


Assuntos
Sequestro de Carbono , Sedimentos Geológicos/química , Plantas Tolerantes a Sal/química , Água do Mar/química , Carbono/análise , Dióxido de Carbono/análise , Grécia , Concentração de Íons de Hidrogênio , Itália , Oceanos e Mares
14.
Mar Environ Res ; 144: 102-110, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30654982

RESUMO

As ocean acidification intensifies, there is growing global concern about the impacts that future pH levels are likely to have on marine life and ecosystems. By analogy, a steep decrease of seawater pH with depth is encountered inside the Kolumbo submarine volcano (northeast Santorini) as a result of natural CO2 venting, making this system ideal for ocean acidification research. Here, we investigated whether the increase of acidity towards deeper layers of Kolumbo crater had any effect on relevant phenotypic traits of bacterial isolates. A total of 31 Pseudomonas strains were isolated from both surface- (SSL) and deep-seawater layers (DSL), with the latter presenting a significantly higher acid tolerance. In particular, the DSL strains were able to cope with H+ levels that were 18 times higher. Similarly, the DSL isolates exhibited a significantly higher tolerance than SSL strains against six commonly used antibiotics and As(III). More importantly, a significant positive correlation was revealed between antibiotics and acid tolerance across the entire set of SSL and DSL isolates. Our findings imply that Pseudomonas species with higher resilience to antibiotics could be favored by the prospect of acidifying oceans. Further studies are required to determine if this feature is universal across marine bacteria and to assess potential ecological impacts.


Assuntos
Ácidos/farmacologia , Antibacterianos/farmacologia , Ecossistema , Fontes Hidrotermais/microbiologia , Pseudomonas/efeitos dos fármacos , Dióxido de Carbono , Concentração de Íons de Hidrogênio , Oceanos e Mares , Pseudomonas/classificação , Pseudomonas/isolamento & purificação , Água do Mar/microbiologia
15.
Environ Health Perspect ; 116(3): 292-6, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18335093

RESUMO

BACKGROUND: The distribution of microorganisms, and especially pathogens, over airborne particles of different sizes has been ignored to a large extent, but it could have significant implications regarding the dispersion of these microorganisms across the planet, thus affecting human health. OBJECTIVES: We examined the microbial quality of the aerosols over the eastern Mediterranean region during an African storm to determine the size distribution of microorganisms in the air. METHODS: We used a five-stage cascade impactor for bioaerosol collection in a coastal city on the eastern Mediterranean Sea during a north African dust storm. Bacterial communities associated with aerosol particles of six different size ranges were characterized following molecular culture-independent methods, regardless of the cell culturability (analysis of 16S rRNA genes). RESULTS: All 16S rDNA clone libraries were diverse, including sequences commonly found in soil and marine ecosystems. Spore-forming bacteria such as Firmicutes dominated large particle sizes (> 3.3 microm), whereas clones affiliated with Actinobacteria (found commonly in soil) and Bacteroidetes (widely distributed in the environment) gradually increased their abundance in aerosol particles of reduced size (< 3.3 microm). A large portion of the clones detected at respiratory particle sizes (< 3.3 microm) were phylogenetic neighbors to human pathogens that have been linked to several diseases. CONCLUSIONS: The presence of aerosolized bacteria in small size particles may have significant implications to human health via intercontinental transportation of pathogens.


Assuntos
Poluentes Atmosféricos/análise , Bactérias/isolamento & purificação , Poeira/análise , África do Norte , Movimentos do Ar , Bactérias/classificação , Bactérias/genética , Infecções Bacterianas/microbiologia , Infecções Bacterianas/prevenção & controle , Infecções Bacterianas/transmissão , Monitoramento Ambiental , Grécia , Humanos , Tamanho da Partícula , RNA Bacteriano/genética , RNA Ribossômico 16S/genética
16.
Environ Sci Pollut Res Int ; 25(24): 23845-23856, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29876858

RESUMO

Soil proteomics, the large-scale characterization of the entire protein complement in soils, provides a promising approach for deciphering the role of microbial functioning in terrestrial ecosystems. However, the extraction of soil proteins in sufficient quantities and of adequate purity remains a challenging task mainly due to the co-extraction of interfering humic substances. Up to now, the treatment of soil extracts with liquid phenol has been the "gold standard" for reducing humics, while the NoviPure cleanup kit was recently launched as a non-toxic approach. The present study describes an alternative method for delivering high-purity proteins based on humic coagulation with trivalent aluminum ions (Al3+). Various experimental parameters were optimized individually in order to maximize protein yield and diminish co-extracted humics. The optimized method was applied on a set of soil samples with diverse physicochemical characteristics and a comparison with the other two techniques was conducted. The amount of residual humics resulting from Al3+-based method was 26 and 35% higher than that from phenol treatment and NoviPure Kit, respectively, but these differences were of marginal statistical significance. With regard to extracted proteins, the average yields of the three methods were comparable, without showing any statistically significant differences. Overall, humic coagulation with Al3+ offers comparable cleanup performance in terms of protein yield and purity, but it is less toxic and less complex than the phenol-partitioning method, whereas it is far less expensive than the NoviPure Kit. The new technique is expected to facilitate the implementation of proteomic studies in soils.


Assuntos
Alumínio/química , Proteínas/isolamento & purificação , Proteômica/métodos , Solo/química , Substâncias Húmicas , Proteínas/análise , Proteínas/química , Microbiologia do Solo
17.
Gigascience ; 6(10): 1-7, 2017 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-29020741

RESUMO

Marine sponges (phylum Porifera) are a diverse, phylogenetically deep-branching clade known for forming intimate partnerships with complex communities of microorganisms. To date, 16S rRNA gene sequencing studies have largely utilised different extraction and amplification methodologies to target the microbial communities of a limited number of sponge species, severely limiting comparative analyses of sponge microbial diversity and structure. Here, we provide an extensive and standardised dataset that will facilitate sponge microbiome comparisons across large spatial, temporal, and environmental scales. Samples from marine sponges (n = 3569 specimens), seawater (n = 370), marine sediments (n = 65) and other environments (n = 29) were collected from different locations across the globe. This dataset incorporates at least 268 different sponge species, including several yet unidentified taxa. The V4 region of the 16S rRNA gene was amplified and sequenced from extracted DNA using standardised procedures. Raw sequences (total of 1.1 billion sequences) were processed and clustered with (i) a standard protocol using QIIME closed-reference picking resulting in 39 543 operational taxonomic units (OTU) at 97% sequence identity, (ii) a de novo clustering using Mothur resulting in 518 246 OTUs, and (iii) a new high-resolution Deblur protocol resulting in 83 908 unique bacterial sequences. Abundance tables, representative sequences, taxonomic classifications, and metadata are provided. This dataset represents a comprehensive resource of sponge-associated microbial communities based on 16S rRNA gene sequences that can be used to address overarching hypotheses regarding host-associated prokaryotes, including host specificity, convergent evolution, environmental drivers of microbiome structure, and the sponge-associated rare biosphere.


Assuntos
Microbiota , Poríferos/microbiologia , Animais , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
18.
Mar Pollut Bull ; 52(11): 1396-405, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16712878

RESUMO

Sediment samples collected from polluted (Augusta Bay) and pristine regions of the Eastern Mediterranean Sea (South Ionian Sea, Thracian Sea) were analyzed for black carbon, aliphatic hydrocarbons and phospholipid ester-linked fatty acids (PLFA). The aim of the study was to investigate the anthropogenic and biogenic inputs into the Eastern Mediterranean Sea and to evaluate the effects of refractory organic matter (e.g. black carbon) and the level of hydrocarbon contamination on benthic microbial community composition. Black carbon, generally considered to be chemically and biologically inert, comprised a significant but highly variable fraction of the sedimentary carbon pool in the analyzed sediments with a ratio to total organic carbon ranging from 0.02 to 0.66. Principal component analysis of the chemical characteristics of the sediments (organic carbon content, black carbon, bioavailable organic carbon, chlorophyll a, phaeopigments, aliphatic hydrocarbons) revealed clustering of samples along a gradient from the most productive and contaminated region of Augusta Bay to the carbon-poor and pristine sediments of the Thracian Sea. PLFA analysis revealed that gram-negative bacteria and microeukaryotes were most abundant in Augusta Bay and in the most impacted station of the Thracian Sea. The high levels of branched and odd-chain fatty acids recorded for these stations is probably linked to the elevated amounts of hydrocarbons at these stations; e.g. microbial communities may have developed the ability to degrade either naturally occurring aliphatic hydrocarbons or hydrocarbons derived from oil contamination.


Assuntos
Bactérias/isolamento & purificação , Carbono/química , Monitoramento Ambiental/estatística & dados numéricos , Poluentes Ambientais/análise , Sedimentos Geológicos/análise , Carbono/análise , Ácidos Graxos/análise , Cromatografia Gasosa-Espectrometria de Massas , Geografia , Hidrocarbonetos/análise , Mar Mediterrâneo , Análise de Componente Principal
19.
Sci Rep ; 6: 28013, 2016 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-27311383

RESUMO

Submarine volcanism represents ~80% of the volcanic activity on Earth and is an important source of mantle-derived gases. These gases are of basic importance for the comprehension of mantle characteristics in areas where subaerial volcanism is missing or strongly modified by the presence of crustal/atmospheric components. Though, the study of submarine volcanism remains a challenge due to their hazardousness and sea-depth. Here, we report (3)He/(4)He measurements in CO2-dominated gases discharged at 500 m below sea level from the high-temperature (~220 °C) hydrothermal system of the Kolumbo submarine volcano (Greece), located 7 km northeast off Santorini Island in the central part of the Hellenic Volcanic Arc (HVA). We highlight that the mantle below Kolumbo and Santorini has a (3)He/(4)He signature of at least 7.0 Ra (being Ra the (3)He/(4)He ratio of atmospheric He equal to 1.39×10(-6)), 3 Ra units higher than actually known for gases-rocks from Santorini. This ratio is also the highest measured across the HVA and is indicative of the direct degassing of a Mid-Ocean-Ridge-Basalts (MORB)-like mantle through lithospheric faults. We finally highlight that the degassing of high-temperature fluids with a MORB-like (3)He/(4)He ratio corroborates a vigorous outgassing of mantle-derived volatiles with potential hazard at the Kolumbo submarine volcano.

20.
Res Microbiol ; 166(5): 448-457, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25908548

RESUMO

The deep eastern basin of the Mediterranean Sea is considered to be one of the world's most oligotrophic areas in the world. Here we performed pyrosequenicng analysis of bacterial and archaeal communities in oxic nutrient-poor sediments collected from the eastern Mediterranean at 1025-4393 m depth. Microbial communities were surveyed by targeting the hypervariable V5-V6 regions of the 16S ribosomal RNA gene using bar-coded pyrosequencing. With a total of 13,194 operational taxonomic units (OTUs) or phylotypes at 97% sequence similarities, the phylogenetic affiliation of microbes was assigned to 23 bacterial and 2 archaeal known phyla, 23 candidate divisions at the phylum level and distributed into 186 families. It was further revealed that the microbial consortia inhabiting all sampling sites were highly diverse, but dominated by phylotypes closely related to members of the genus Pseudomonas and Marine Group I archaea. Such pronounced and widespread enrichment probably manifests the cosmopolitan character of these species and raises questions about their metabolic adaptation to the physical stressors and low nutrient availability of the deep eastern Mediterranean Sea.


Assuntos
Archaea/classificação , Archaea/isolamento & purificação , Bactérias/classificação , Bactérias/isolamento & purificação , Sedimentos Geológicos/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala , Consórcios Microbianos , Archaea/genética , Archaea/metabolismo , Bactérias/genética , Bactérias/metabolismo , Biodiversidade , DNA Arqueal/genética , DNA Bacteriano/genética , Mar Mediterrâneo , Filogenia , Pseudomonas/classificação , Pseudomonas/genética , Pseudomonas/isolamento & purificação , RNA Ribossômico 16S/genética , Água do Mar/microbiologia , Análise de Sequência de DNA
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