Search

Salt water, in addition to being a naturally corrosive environment, also includes factors such as temperature, pressure, and the presence of the microbial community in the environment that influence degradation processes on metal surfaces. The presence or absence of water flow over the metal surfaces is also an important aspect that influences the corrosion of metals. The objective of this study was to evaluate the presence or absence of salt water flow in the formation of biofilms grown in 316L stainless steel coupons. For this, the 316L stainless steel coupons were exposed in two different microcosms, the first being a system with continuous salt water flow, and the second without salt water flow system. The results of the sequencing of the 16S rDNA genes showed a clear difference in structures and diversity between the evaluated biofilms. There was greater abundance and diversity in the “In Flux” system when compared to the “No Flux” biofilm. The analysis of bacterial diversity showed a predominance of the Gammaproteobacteria class in both systems. However, at lower taxonomic levels, there were considerable differences in representativeness. Representatives of Vibrionales, Alteromonadales, Oceanospirillales, and Flavobacteriales were predominant in “No Flux”, whereas in “In Flux” there was a greater representation of Alteromonadales, Rhodobacterales, and Saprospirales. These findings help to understand how the flow of water influences the dynamics of the formation of microbial biofilms on metal surfaces, which will contribute to the choice of strategies used to mitigate microbial biofouling.

As much as 400 Tg of carbon from airborne semivolatile aromatic hydrocarbons is deposited to the oceans every year, the largest identified source of anthropogenic organic carbon to the ocean. Microbial degradation is a key sink of these pollutants in surface waters, but has received little attention in polar environments. We have challenged Antarctic microbial communities from the sea-surface microlayer (SML) and the subsurface layer (SSL) with polycyclic aromatic hydrocarbons (PAHs) at environmentally relevant concentrations. PAH degradation rates and the microbial responses at both taxonomical and functional levels were assessed. Evidence for faster removal rates was observed in the SML, with rates 2.6-fold higher than in the SSL. In the SML, the highest removal rates were observed for the more hydrophobic and particle-bound PAHs. After 24 h of PAHs exposure, particle-associated bacteria in the SML showed the highest number of significant changes in their composition. These included significant enrichments of several hydrocarbonoclastic bacteria, especially the fast-growing genera Pseudoalteromonas, which increased their relative abundances by eightfold. Simultaneous metatranscriptomic analysis showed that the free-living fraction of SML was the most active fraction, especially for members of the order Alteromonadales, which includes Pseudoalteromonas. Their key role in PAHs biodegradation in polar environments should be elucidated in further studies. This study highlights the relevant role of bacterial populations inhabiting the sea-surface microlayer, especially the particle-associated habitat, as relevant bioreactors for the removal of aromatic hydrocarbons in the oceans., We thank the staff of the Marine Technology Unit (UTM-CSIC) for their logistical support during the sampling campaign at Livingston Island. We also thank Dr. Daniel Lundin for unconditional bioinformatics support. Special thanks to TERNUA for their non-profit collaboration by sponsoring with technical ecofriendly clothing and gear equipment for Antarctic campaigns and for increasing our project visibility.

The acidification and warming of seawater have several impacts on marine organisms, including over microorganisms. The influence of acidification and warming of seawater on biofilms grown on API 5L steel surfaces was evaluated by sequencing the 16S ribosomal gene. For this, three microcosms were designed, the first simulating the natural marine environment (MCC), the second with a decrease in pH from 8.1 to 7.9, and an increase in temperature by 2 °C (MMS), and the third with pH in around 7.7 and an increase in temperature of 4 °C (MES). The results showed that MCC was dominated by the Gammaproteobacteria class, mainly members of the Alteromonadales Order. The second most abundant group was Alphaproteobacteria, with a predominance of Rhodobacterales and Oceanospirillales. In the MMS system there was a balance between representatives of the Gammaproteobacteria and Alphaproteobacteria classes. In MES there was an inversion in the representations of the most prevalent classes previously described in MCC. In this condition, there was a predominance of members of the Alphaproteobacteria Class, in contrast to the decrease in the abundance of Gammaproteobacteria members. These results suggest that possible future climate changes may influence the dynamics of the biofouling process in surface metals. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00925-7.

It is now indisputable that plastics are ubiquitous and problematic in ecosystems globally. Many suggestions have been made about the role that biofilms colonizing plastics in the environment—termed the “Plastisphere”—may play in the transportation and ecological impact of these plastics. By collecting and re-analyzing all raw 16S rRNA gene sequencing and metadata from 2,229 samples within 35 studies, we have performed the first meta-analysis of the Plastisphere in marine, freshwater, other aquatic (e.g., brackish or aquaculture) and terrestrial environments. We show that random forest models can be trained to differentiate between groupings of environmental factors as well as aspects of study design, but—crucially—also between plastics when compared with control biofilms and between different plastic types and community successional stages. Our meta-analysis confirms that potentially biodegrading Plastisphere members, the hydrocarbonoclastic Oceanospirillales and Alteromonadales are consistently more abundant in plastic than control biofilm samples across multiple studies and environments. This indicates the predilection of these organisms for plastics and confirms the urgent need for their ability to biodegrade plastics to be comprehensively tested. We also identified key knowledge gaps that should be addressed by future studies.

Despite the widespread distribution of proteorhodopsin (PR)-containing bacteria in the oceans, the use of light-derived energy to promote bacterial growth has only been shown in a few bacterial isolates, and there is a paucity of data describing the metabolic effects of light on environmental photoheterotrophic taxa. Here, we assessed the effects of light on the taxonomic composition, cell integrity and growth responses of microbial communities in monthly incubations between spring and autumn under different environmental conditions. The photoheterotrophs expressing PR in situ were dominated by Pelagibacterales and SAR116 in July and November, while members of Euryarchaeota, Gammaproteobacteria and Bacteroidetes dominated the PR expression in spring. Cell-membrane integrity decreased under dark conditions throughout most of the assessment, with maximal effects in summer, under low-nutrient conditions. A positive effect of light on growth was observed in one incubation (out of nine), coinciding with a declining phytoplankton bloom. Light-enhanced growth was found in Gammaproteobacteria (Alteromonadales) and Bacteroidetes (Polaribacter and Tenacibaculum). Unexpectedly, some Pelagibacterales also exhibited higher growth rates under light conditions. We propose that the energy harvested by PRs helps to maintain cell viability in dominant coastal photoheterotrophic oligotrophs while promoting the growth of some widespread taxa benefiting from the decline of phytoplankton blooms.

International audience; Microbial anodes and oxygen reducing microbial cathodes were designed separately under constant polarization at +0.1 V/SCE in a hypersaline medium (NaCl 45 g/L). They were then associated to design two-compartment microbial fuel cells (MFCs). These MFCs produced up to 209 ±24 mW m-2 during a week. This was the first demonstration that hypersaline MFCs equipped with microbial cathodes can produce power density at this level. Desulfuromonas sp. were confirmed to be key species of the anodes. The efficiency of the cathodes was linked to the development of a redox system centred at +0.2 V/SCE and to the presence of Gammaproteobacteria (Alteromonadales and Oceanospirillales), especially an unclassified order phylogenetically linked to the genus Thioalobacter. Comparing the different performance of the four MFCs with the population analyses suggested that polarization at +0.1V/SCE should be maintained longer to promote the growth of Thioalobacter on the cathode and thus increase the MFC performance.

Bacterial communities colonized on submerged substrata are recognized as a key factor in the formation of complex biofouling phenomenon in the marine environment. Despite massive maritime activities and a large industrial sector in the nearshore of the Laccadive Sea, studies describing pioneer bacterial colonizers and community succession during the early-stage biofilm are scarce. We investigated the biofilm-forming bacterial community succession on three substrata viz. stainless steel, high-density polyethylene, and titanium over 15 days of immersion in the seawater intake area of a power plant, located in the southern coastal region of India. The bacterial community composition of biofilms and peripheral seawater were analyzed by Illumina MiSeq sequenced 16S rRNA gene amplicons. The obtained metataxonomic results indicated a profound influence of temporal succession over substrate type on the early-stage biofilm-forming microbiota. Bacterial communities showed vivid temporal dynamics that involved variations in abundant bacterial groups. The proportion of dominant phyla viz. Proteobacteria decreased over biofilm succession days, while Bacteroidetes increased, suggesting their role as initial and late colonizers, respectively. A rapid fluctuation in the proportion of two bacterial orders viz. Alteromonadales and Vibrionales were observed throughout the successional stages. LEfSe analysis identified specific bacterial groups at all stages of biofilm development, whereas no substrata type-specific groups were observed. Furthermore, the results of PCoA and UPGMA hierarchical clustering demonstrated that the biofilm-forming community varied considerably from the planktonic community. Phylum Proteobacteria preponderated the biofilm-forming community, while the Bacteroidetes, Cyanobacteria, and Actinobacteria dominated the planktonic community. Overall, our results refute the common assumption that substrate material has a decisive impact on biofilm formation; rather, it portrayed that the temporal succession overshadowed the influence of the substrate material. Our findings provide a scientific understanding of the factors shaping initial biofilm development in the marine environment and will help in designing efficient site-specific anti-biofouling strategies.

Polyphasic taxonomic analysis was performed to characterize a novel bacterium, which was isolated from surface sediment of the Terra Nova Bay, Antarctica, and designated as R04H25T. The cells of the isolate were Gram-stain-negative, aerobic, non-motile, slightly curved rods. Growth occurred at 4–42 °C, pH 7.0–9.5, and in 1–15 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences indicated that strain R04H25T formed an independent lineage within the genus Pseudidiomarina and its nearest neighbours were Pseudidiomarina donghaiensis 908033T (98.2 %), Pseudidiomarina marina PIM1T (98.1 %), Pseudidiomarina woesei W11T (97.8 %), Pseudidiomarina maritima 908087T (97.1 %) and Pseudidiomarina tainanensis PIN1T (97.0 %). The average nucleotide identities between strain R04H25T and the nearest neighbours were 76.2–77.7 %. The major fatty acids were iso-C17 : 0, summed feature 9, iso-C15 : 0, C16 : 0 and iso-C11 : 0 3-OH. The polar lipids comprised phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminophospholipid, three unidentified glycolipids and two unidentified lipids. The predominant respiratory quinone was ubiquinone 8. The genomic DNA G+C content was 48.2 mol%. On the basis of the phylogenetic, physiological and chemotaxonomic results, we propose a novel species named as Pseudidiomarina gelatinasegens sp. nov. in the genus Pseudidiomarina , with the type strain R04H25T (=GDMCC 1.1503T=KCTC 62911T).

The main studies on the corrosion of metals induced by microorganisms are directed only to the surface of the metal, without considering the presence of welds between these surfaces. For this reason, we evaluated the difference of microbial community grown in carbon steel coupons, and two different types of welds, E7018 and Tungsten electrodes, exposed under simulated microcosm. After 30 days, they were recovered, the biofilms scraped and the microbial communities analyzed by 16S rRNA gene sequencing. The results showed that there was a differentiated distribution among the three samples collected. Proteobacteria phylum composed most of the species described in all samples. At the class level, Gammaproteobacteria was the most detected, followed by Alphaproteobacteria and Flavobacteriia. The most prevalent order was Alteromonadales, which was present in Weld2, followed by Rhodobacteriales, which was more prevalent in Fe1020 and Weld1. The orders Cytophagales, Sphingomonadales, and Burkholderiales were described in higher number in Fe1020, whereas Oceanospirillales, Thiotrichales, Flavobacteriales, Rhodospirillales, and Kordiimonadales were higher in samples Weld1 and Weld2. The analyses between the three evaluated conditions show the presences of bacterial groups preferred by different types of metal, suggesting that approaches in the control of biocorrosion should take into account the chemical composition of the metal.

Protease-producing bacteria and their extracellular proteases are key players in degrading organic nitrogen to drive marine nitrogen cycling and yet knowledge on both of them is still very limited. This study screened protease-producing bacteria from the South China Sea sediments and analyzed the diversity of their extracellular proteases at the family level through N-terminal amino acid sequencing. Results of the 16S rRNA gene sequence analysis showed that all screened protease-producing bacteria belonged to the class Gammaproteobacteria and most of them were affiliated with different genera within the orders Alteromonadales and Vibrionales. The N-terminal amino acid sequence analysis for fourteen extracellular proteases from fourteen screened bacterial strains revealed that all these proteases belonged to the M4 family of metalloproteases or the S8 family of serine proteases. This study presents new details on taxa of marine sedimentary protease-producing bacteria and types of their extracellular proteases, which will help to comprehensively understand the process and mechanism of the microbial enzymatic degradation of marine sedimentary organic nitrogen.

Global climate change is significantly affecting marine life off the northern tip of the Antarctic Peninsula, but little is known about microbial ecology in this area. The main goal of this study was to investigate the bacterioplankton community structure in surface waters using pyrosequencing and to determine factors influencing this community. Pelagibacterales and Rhodobacterales (Alphaproteobacteria), Oceanospirillales and Alteromonadales (Gammaproteobacteria), and Flavobacteriales (Bacteroidetes) were the core taxa in our samples, and the five most relatively abundant genera were Pelagibacter, Polaribacter, Octadecabacter, group HTCC2207 and Sulfitobacter. Although nutrients and chlorophyll a (chl a) contributed more to bacterioplankton community structure than water masses or depth, only 30.39% of the variance could be explained by the investigated environmental factors, as revealed by RDA and pRDA. No significant difference with respect to nutrients and chl a was observed among water masses or depth, as indicated by ANOVA. Furthermore, significant correlations among the dominant bacterial genera were more common than correlations between dominant genera and environmental factors, as revealed by Spearman analysis. We conclude that nutrients and chl a become homogeneous and that interpopulation interactions may have a central role in influencing the bacterial community structure in surface waters off the northern tip of the Antarctic Peninsula during the summer.

The twilight zone (from the base of the euphotic zone to the depth of 1,000 m) is the major area of particulate organic carbon (POC) remineralization in the ocean, and heterotrophic microbes contribute to more than 70% of the estimated remineralization. However, little is known about the microbial community and metabolic activity directly associated with POC remineralization in this chronically understudied realm. Here, we characterized the microbial community proteomes of POC samples collected from the twilight zone of three contrasting sites in the Northwest Pacific Ocean using a metaproteomic approach. The particle-attached bacteria from Alteromonadales, Rhodobacterales, and Enterobacterales were the primary POC remineralizers. Hydrolytic enzymes, including proteases and hydrolases, that degrade proteinaceous components and polysaccharides, the main constituents of POC, were abundant and taxonomically associated with these bacterial groups. Furthermore, identification of diverse species-specific transporters and metabolic enzymes implied niche specialization for nutrient acquisition among these bacterial groups. Temperature was the main environmental factor driving the active bacterial groups and metabolic processes, and Enterobacterales replaced Alteromonadales as the predominant group under low temperature. This study provides insight into the key bacteria and metabolic processes involved in POC remineralization, and niche complementarity and species substitution among bacterial groups are critical for efficient POC remineralization in the twilight zone. IMPORTANCE The ocean's twilight zone is a critical zone where more than 70% of the sinking particulate organic carbon (POC) is remineralized. Therefore, the twilight zone determines the size of biological carbon storage in the ocean and regulates the global climate. Prokaryotes are major players that govern remineralization of POC in this region. However, knowledge of microbial community structure and metabolic activity is still lacking. This study unveiled microbial communities and metabolic activities of POC samples collected from the twilight zone of three contrasting environments in the Northwest Pacific Ocean using a metaproteomic approach. Alteromonadales, Rhodobacterales, and Enterobacterales were the major remineralizers of POC. They excreted diverse species-specific hydrolytic enzymes to split POC into solubilized POC or dissolved organic carbon. Temperature played a crucial role in regulating the community composition and metabolism. Furthermore, niche complementarity or species substitution among bacterial groups guaranteed the efficient remineralization of POC in the twilight zone.

Background In octocorals (Cnidaria Octocorallia), the functional relationship between host health and its symbiotic consortium has yet to be determined. Here, we employed comparative metagenomics to uncover the distinct functional and phylogenetic features of the microbiomes of healthy Eunicella gazella, Eunicella verrucosa, and Leptogorgia sarmentosa tissues, in contrast with the microbiomes found in seawater and sediments. We further explored how the octocoral microbiome shifts to a pathobiome state in E. gazella. Results Multivariate analyses based on 16S rRNA genes, Clusters of Orthologous Groups of proteins (COGs), Protein families (Pfams), and secondary metabolite-biosynthetic gene clusters annotated from 20 Illumina-sequenced metagenomes each revealed separate clustering of the prokaryotic communities of healthy tissue samples of the three octocoral species from those of necrotic E. gazella tissue and surrounding environments. While the healthy octocoral microbiome was distinguished by so-far uncultivated Endozoicomonadaceae, Oceanospirillales, and Alteromonadales phylotypes in all host species, a pronounced increase of Flavobacteriaceae and Alphaproteobacteria, originating from seawater, was observed in necrotic E. gazella tissue. Increased abundances of eukaryotic-like proteins, exonucleases, restriction endonucleases, CRISPR/Cas proteins, and genes encoding for heat-shock proteins, inorganic ion transport, and iron storage distinguished the prokaryotic communities of healthy octocoral tissue regardless of the host species. An increase of arginase and nitric oxide reductase genes, observed in necrotic E. gazella tissues, suggests the existence of a mechanism for suppression of nitrite oxide production by which octocoral pathogens may overcome the host’s immune system. Conclusions This is the first study to employ primer-less, shotgun metagenome sequencing to unveil the taxonomic, functional, and secondary metabolism features of prokaryotic communities in octocorals. Our analyses reveal that the octocoral microbiome is distinct from those of the environmental surroundings, is host genus (but not species) specific, and undergoes large, complex structural changes in the transition to the dysbiotic state. Host-symbiont recognition, abiotic-stress response, micronutrient acquisition, and an antiviral defense arsenal comprising multiple restriction endonucleases, CRISPR/Cas systems, and phage lysogenization regulators are signatures of prokaryotic communities in octocorals. We argue that these features collectively contribute to the stabilization of symbiosis in the octocoral holobiont and constitute beneficial traits that can guide future studies on coral reef conservation and microbiome therapy.

The obstacle to optimal utilization of biogas technology is poor understanding of biogas microbiomes diversities over a wide geographical coverage. We performed random shotgun sequencing on twelve environmental samples. Randomized complete block design was utilized to assign the twelve treatments to four blocks, within eastern and central regions of Kenya. We obtained 42 million paired-end reads that were annotated against sixteen reference databases using two ENVO ontologies, prior to β-diversity studies. We identified 37 phyla, 65 classes and 132 orders. Bacteria dominated and comprised 28 phyla, 42 classes and 92 orders, conveying substrate’s versatility in the treatments. Though, Fungi and Archaea comprised 5 phyla, the Fungi were richer; suggesting the importance of hydrolysis and fermentation in biogas production. High β-diversity within the taxa was largely linked to communities’ metabolic capabilities. Clostridiales and Bacteroidales, the most prevalent guilds, metabolize organic macromolecules. The identified Cytophagales, Alteromonadales, Flavobacteriales, Fusobacteriales, Deferribacterales, Elusimicrobiales, Chlamydiales, Synergistales to mention but few, also catabolize macromolecules into smaller substrates to conserve energy. Furthermore, δ-Proteobacteria, Gloeobacteria and Clostridia affiliates syntrophically regulate PH2 and reduce metal to provide reducing equivalents. Methanomicrobiales and other Methanomicrobia species were the most prevalence Archaea, converting formate, CO2(g), acetate and methylated substrates into CH4(g). Thermococci, Thermoplasmata and Thermoprotei were among the sulfur and other metal reducing Archaea that contributed to redox balancing and other metabolism within treatments. Eukaryotes, mainly fungi were the least abundant guild, comprising largely Ascomycota and Basidiomycota species. Chytridiomycetes, Blastocladiomycetes and Mortierellomycetes were among the rare species, suggesting their metabolic and substrates limitations. Generally, we observed that environmental and treatment perturbations influenced communities’ abundance, β-diversity and reactor performance largely through stochastic effect. Understanding diversity of biogas microbiomes over wide environmental variables and its’ productivity provided insights into better management strategies that ameliorate biochemical limitations to effective biogas production.

Although several hypotheses have been proposed, the significance of secondary genetic elements remains uncertain. In an effort to extend our current knowledge on multipartite genomes we studied how different gene categories are distributed and expressed in Pseudoalteromonas. Our data support that secondary replicons are open and flexible in nature, and likely serve a variety of specialized roles in bacteria. This work also adds to the growing body of evidence demonstrating that bacterial cells are highly organized at all levels., Bacterial genomes typically consist of one large chromosome, but can also include secondary replicons. These so-called multipartite genomes are scattered on the bacterial tree of life with the majority of cases belonging to Proteobacteria. Within the class gamma-proteobacteria, multipartite genomes are restricted to the two families Vibrionaceae and Pseudoalteromonadaceae. Whereas the genome of vibrios is well studied, information on the Pseudoalteromonadaceae genome is much scarcer. We have studied Pseudoalteromonadaceae with respect to the origin of the chromid, how pangene categories are distributed, how genes are expressed relative to their genomic location, and identified chromid hallmark genes. We calculated the Pseudoalteromonadaceae pangenome based on 25 complete genomes and found that core/softcore are significantly overrepresented in late replicating sectors of the chromid, regardless of how the chromid is replicated. On the chromosome, core/softcore and shell/cloud genes are only weakly overrepresented at the chromosomal replication origin and termination sequences, respectively. Gene expression is trending downwards with increasing distance from the chromosomal oriC, whereas the chromidal expression pattern is more complex. Moreover, we identified 78 chromid hallmark genes, and BLASTp searches suggest that the majority of them were acquired from the ancestral gene pool of Alteromonadales. Finally, our data strongly suggest that the chromid originates from a plasmid that was acquired in a relatively recent event. In summary, this study extends our knowledge on multipartite genomes, and helps us understand how and why secondary replicons are acquired, why they are maintained, and how they are shaped by evolution.

Summary Elucidating the mechanisms controlling the synthesis of hydroxyectoine is important to design novel genetic engineering strategies for optimizing the production of this biotechnologically relevant compatible solute. The genome of the halophilic bacterium Chromohalobacter salexigens carries two ectoine hydroxylase genes, namely ectD and ectE, whose encoded proteins share the characteristic consensus motif of ectoine hydroxylases but showed only a 51.9% identity between them. In this work, we have shown that ectE encodes a secondary functional ectoine hydroxylase and that the hydroxyectoine synthesis mediated by this enzyme contributes to C.␣salexigens thermoprotection. The evolutionary pattern of EctD and EctE and related proteins suggests that they may have arisen from duplication of an ancestral gene preceding the directional divergence that gave origin to the orders Oceanospirillales and Alteromonadales. Osmoregulated expression of ectD at exponential phase, as well as the thermoregulated expression of ectD at the stationary phase, seemed to be dependent on the general stress factor RpoS. In contrast, expression of ectE was always RpoS‐dependent regardless of the growth phase and osmotic or heat stress conditions tested. The data presented here suggest that the AraC‐GlxA‐like EctZ transcriptional regulator, whose encoding gene lies upstream of ectD, plays a dual function under exponential growth as both a transcriptional activator of osmoregulated ectD expression and a repressor of ectE transcription, privileging the synthesis of the main ectoine hydroxylase EctD. Inactivation of ectZ resulted in a higher amount of the total ectoines pool at the expenses of a higher accumulation of ectoine, with maintenance of the hydroxyectoine levels. In addition to the transcriptional control, our results suggest a strong post‐transcriptional regulation of hydroxyectoine synthesis. Data on the accumulation of ectoine and hydroxyectoine in rpoS and ectZ strains pave the way for using these genetic backgrounds for metabolic engineering for hydroxyectoine production., Besides the main ectoine hydroxylase EctD, the halophilic bacterium Chromohalobacter salexigens possess a secondary ectoine hydroxylases, EctE, which contributes to C. salexigens thermoprotection. Expression of ectD and ectE is dependent on the growth phase and the general stress factor RpoS. In addition, the EctZ transcriptional regulator plays a dual function under exponential growth as both an activator of osmorregulated ectD expression and a repressor of ectE transcription, privileging the synthesis of the main ectoine hydroxylase EctD.

Algal polysaccharides constitute a diverse and abundant reservoir of organic matter for marine heterotrophic bacteria, central to the oceanic carbon cycle. We investigated the uptake of alginate, a major brown macroalgal polysaccharide, by microbial communities from kelp-dominated coastal habitats. Congruent with cell growth and rapid substrate utilization, alginate amendments induced a decrease in bacterial diversity and a marked compositional shift towards copiotrophic bacteria. We traced 13C derived from alginate into specific bacterial incorporators and quantified the uptake activity at the single-cell level, using halogen in situ hybridization coupled to nanoscale secondary ion mass spectrometry (HISH-SIMS) and DNA stable isotope probing (DNA-SIP). Cell-specific alginate uptake was observed for Gammaproteobacteria and Flavobacteriales, with carbon assimilation rates ranging from 0.14 to 27.50 fg C µm−3 h−1. DNA-SIP revealed that only a few initially rare Flavobacteriaceae and Alteromonadales taxa incorporated 13C from alginate into their biomass, accounting for most of the carbon assimilation based on bulk isotopic measurements. Functional screening of metagenomic libraries gave insights into the genes of alginolytic Alteromonadales active in situ. These results highlight the high degree of niche specialization in heterotrophic communities and help constraining the quantitative role of polysaccharide-degrading bacteria in coastal ecosystems.

The performance of a commercial starter culture was investigated in a new marine post-smolt RAS, analyzing the microbial communities of 100 samples collected monthly over a year from biofilter biofilm carriers, tank wall biofilm, production water and fish skin. Totally 139 operational taxonomic units (OTUs) were defined in the starter culture, of which the classified members of Rhodobacterales, Bacteroidetes, Alteromonadales and Planctomycetes were largely the first colonizers of the biofilter carriers. Early colonizing OTUs that dominated biofilter biofilm carriers (> 5% relative abundance) were stably present over time, but the development went slowly from a few OTUs with very high relative abundance to several dominant ones with lower relative abundance. Operating taxonomic units not associated with the starting culture became prominent on the biofilter biofilm carriers only towards the end of the trial period. These were termed environmental OTUs. Comparing the two OTU quantitives in a ratio, where counts were based on all OTUs in the sample, the starter culture OTUs:environmental OTUs were 1.2 and 0.9 at the first and last sampling time for the biofilter biofilm carriers. Correspondingly, for all defined OTUs in the RAS sampling sites together, the ratio changed from 0.8 to 0.6 during experiment. Independent of origin, omniscient OTUs at a sampling site, did also have the highest relative abundances and were normally shared between biofilter biofilm carriers and the production water. New and lost OTUs between sampling times were on average 44 % of the OTUs defined, and this OTUflow was strongest for low abundant environmental OTUs. The maturation of the biofilter with respect to nitrification took long time, and the Nitrospira strain in the starter culture was not adapted to marine salinities. Still, we report a controlled colonization of the marine RAS by the starter culture. publishedVersion

The Persian Gulf, hosting ca. 48% of the world’s oil reserves, has been chronically exposed to natural oil seepage. Oil spill studies show a shift in microbial community composition in response to oil pollution; however, the influence of chronic oil exposure on the microbial community remains unknown. We performed genome-resolved comparative analyses of the water and sediment samples along Persian Gulf’s pollution continuum (Strait of Hormuz, Asalouyeh, and Khark Island). Continuous exposure to trace amounts of pollution primed the intrinsic and rare marine oil-degrading microbes such as Oceanospirillales, Flavobacteriales, Alteromonadales, and Rhodobacterales to bloom in response to oil pollution in Asalouyeh and Khark samples. Comparative analysis of the Persian Gulf samples with 106 oil-polluted marine samples reveals that the hydrocarbon type, exposure time, and sediment depth are the main determinants of microbial response to pollution. High aliphatic content of the pollution enriched for Oceanospirillales, Alteromonadales, and Pseudomonadales whereas, Alteromonadales, Cellvibrionales, Flavobacteriales, and Rhodobacterales dominate polyaromatic polluted samples. In chronic exposure and oil spill events, the community composition converges towards higher dominance of oil-degrading constituents while promoting the division of labor for successful bioremediation.

14 pages, 4 figures, 1 table.-- This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY), To study the response of bacteria to different size-fractions of naturally occurring dissolved organic matter (DOM), a natural prokaryotic community from North Atlantic mesopelagic waters (1000 m depth) was isolated and grown in (i) 0.1-μm filtered seawater (CONTROL), (ii) the low-molecular-weight (1 kDa) and low-molecular-weight DOM fractions (H + L-DOM), to test the potential effect of ultrafiltration on breaking the DOM size continuum. Prokaryotic abundance and leucine incorporation were consistently higher in the H + L-DOM niche than in the L-DOM and CONTROL treatments, suggesting a different interaction with each DOM fraction and the disruption of the structural DOM continuum by ultrafiltration, respectively. Rhodobacterales (Alphaproteobacteria) and Flavobacteriales (Bacteroidetes) were particularly enriched in L-DOM and closely related to the colored DOM (CDOM) fraction, indicating the tight link between these groups and changes in DOM aromaticity. Conversely, some other taxa that were rare or undetectable in the original bacterial community were enriched in the H + L-DOM treatment (e.g., Alteromonadales belonging to Gammaproteobacteria), highlighting the role of the rare biosphere as a seed bank of diversity against ecosystem disturbance. The relationship between the fluorescence of protein-like CDOM and community composition of populations in the H + L-DOM treatment suggested their preference for labile DOM. Conversely, the communities growing on the L-DOM niche were coupled to humic-like CDOM, which may indicate their ability to degrade more reworked DOM and/or the generation of refractory substrates (as by-products of the respiration processes). Most importantly, L- and/or H + L-DOM treatments stimulated the growth of unique bacterial amplicon sequence variants (ASVs), suggesting the potential of environmental selection (i.e., changes in DOM composition and availability), particularly in the light of climate change scenarios. Taken together, our results suggest that different size-fractions of DOM induced niche-specialization and differentiation of mesopelagic bacterial communities, Funding for sampling and analyses was supported by the projects “Biodiversidade Funcional do Microplancton nas profundidades mariñas de Galicia” (BIO-PROF, Ref. 10MMA604024PR, 2010–2012, Xunta de Galicia), “Fuentes de Materia Orgánica y Diversidad Funcional del Microplancton en las aguas profundas del Atlántico Norte” (MODUPLAN, Ref. CTM 2011-24008, 2012–2015, Plan Nacional I + D + I; https://moduplansarmiento2014.wordpress.com/) to MMV, and “Deep Standard Oceanographic Sections monitoring program RADIALES-PRODUNDOS”, funded by the Instituto Español de Oceanografía (IEO). Additional funding was provided by the Axencia Galega de Innovación (GAIN, Xunta de Galicia) through IEO-GAIN Programme Contracts (Contratos Programa). TR-R was supported by MICINN program “Personal Técnico de Apoyo a la Investigación” (PTA, Ref. PTA2015-10948-I). MN-C was funded by the projects HOTMIX and FERMIO (MINECO, CTM2011–30010–C02–MAR and CTM2014-57334-JIN, respectively), both co-financed with FEDER funds. EG-F was supported by the BIO-PROF and MODUPLAN projects

Orders Oceanospirillales and Pseudomonadales play important roles in various ecosystems as the keystone taxa of microbiomes. However, the two orders present a close evolutionary relationship, which might have caused taxonomic misinterpretation and resulted in an incorrect understanding of their evolutionary history. In this study, first, we used the 16S rRNA gene sequences of 2,049 species of Gammaproteobacteria to build a phylogenetic tree, which demonstrated that reports regarding the evolutionary relationship of orders Cellvibrionales, Oceanospirillales, and Pseudomonadales based on a single conserved gene with a poor resolution have been conflicting; in particular, the major families Moraxellaceae and Pseudomonadaceae of order Pseudomonadales were separated from orders Cellvibrionales and Oceanospirillales. Subsequently, we constructed the bac120 trees of all representative reference genomes of class Gammaproteobacteria based on 120 ubiquitous single-copy proteins from bacteria and a phylogenomic tree based on the 119 core genes of 257 reference genomes obtained from orders Cellvibrionales, Oceanospirillales, and Pseudomonadales to cross validate and infer their intrinsic evolutionary relationships. These results indicated that two novel orders, Moraxellales ord. nov. and Kangiellales ord. nov., and three novel families, Marinobacteraceae fam. nov., Perlucidibacaceae fam. nov., and Zooshikellaceae fam. nov., should be proposed. Additionally, orders Cellvibrionales and Oceanospirillales were merged into the order Pseudomonadales except for families Moraxellaceae and Kangiellaceae in class Gammaproteobacteria, which currently includes 18 families. Our work sheds some light on the evolutionary history of class Gammaproteobacteria, which could facilitate the detection and taxonomic analysis of natural communities. IMPORTANCE The orders Cellvibrionales, Oceanospirillales, and Pseudomonadales, as three major orders of the largest bacterial class, Gammaproteobacteria, play important roles in various ecosystems as the keystone taxa of microbiomes, but their evolutionary relationship is currently polyphyletic and chaotic. Here, we constructed a bac120 tree and core-genome tree and calculated the amino acid identity (AAI) value to explore their intrinsic evolutionary history. In this study, we proposed two novel orders and three novel families. This evolution study vastly reconstructed the taxonomic framework of class Gammaproteobacteria and could provide a more distinct perspective on global distribution and evolutionary patterns of these environmental microorganisms.

International audience; Photosynthetic microbial mats are stable, self-supported communities. Due to their coastal localization, these mats are frequently exposed to hydrocarbon contamination and are able to grow on it. To decipher how this contamination disturbs the functioning of microbial mats, we compared two mats: a contaminated mat exposed to chronic petroleum contamination and a reference mat. The taxonomic and metabolic structures of the mats in spring and fall were determined using metagenomic and metatranscriptomic approaches. Extremely high contamination disturbed the seasonal variations of the mat. ABC transporters, two-component systems, and type IV secretion system-related genes were overabundant in the contaminated mats. Xenobiotic degradation metabolism was minor in the metagenomes of both mats, and only the expression of genes involved in polycyclic aromatic hydrocarbon degradation was higher in the contaminated mat. Interestingly, the expression rates of genes involved in hydrocarbon activation decreased during the 1-year study period, concomitant with the decrease in easily degradable hydrocarbons, suggesting a transient effect of hydrocarbon contamination. Alteromonadales and Oceanospirillales hydrocarbonoclastic bacteria appeared to be key in hydrocarbon remediation in the contaminated mat. Overall, the contaminated microbial mat was able to cope with hydrocarbon contamination and displayed an adaptive functioning that modified seasonal behaviour.

The large ribosomal RNAs of eukaryotes frequently contain expansion sequences that add to the size of the rRNAs but do not affect their overall structural layout and are compatible with major ribosomal function as an mRNA translation machine. The expansion of prokaryotic ribosomal RNAs is much less explored. In order to obtain more insight into the structural variability of these conserved molecules, we herein report the results of a comprehensive search for the expansion sequences in prokaryotic 5S rRNAs. Overall, 89 expanded 5S rRNAs of 15 structural types were identified in 15 archaeal and 36 bacterial genomes. Expansion segments ranging in length from 13 to 109 residues were found to be distributed among 17 insertion sites. The strains harboring the expanded 5S rRNAs belong to the bacterial orders Clostridiales, Halanaerobiales, Thermoanaerobacterales, and Alteromonadales as well as the archael order Halobacterales. When several copies of 5S rRNA gene are present in a genome, the expanded versions may co-exist with normal 5S rRNA genes. The insertion sequences are typically capable of forming extended helices, which do not seemingly interfere with folding of the conserved core. The expanded 5S rRNAs have largely been overlooked in 5S rRNA databases.

Halophiles are relatively unexplored as potential sources of novel species. However, little is known about the culturable bacterial diversity thrive in hypersaline lakes. In this work, a total of 343 bacteria from sediment samples of Aiding Lake, China, were isolated using nine different media supplemented with 5% or 15% (w/v) NaCl. The number of species and genera of bacteria recovered from the different media varied, indicating the need to optimize the isolation conditions. The results showed an unexpected level of bacterial diversity, with four phyla (Actinobacteria, Firmicutes, Proteobacteria, and Rhodothermaeota), fourteen orders (Actinopolysporales, Alteromonadales, Bacillales, Balneolales, Chromatiales, Glycomycetales, Jiangellales, Micrococcales, Micromonosporales, Oceanospirillales, Pseudonocardiales, Rhizobiales, Streptomycetales, and Streptosporangiales), including 17 families, 43 genera (including two novel genera), and 71 species (including four novel species). The predominant phyla included Actinobacteria and Firmicutes and the predominant genera included Actinopolyspora, Gracilibacillus, Halomonas, Nocardiopsis, and Streptomyces. To our knowledge, this is the first time that members of phylum Rhodothermaeota were identified in sediment samples from a salt lake.

The obstacle to optimal utilization of biogas technology is poor understanding of biogas’ microbiome diversities over a wide geographical coverage. We performed random shotgun sequencing on twelve environmental samples. A randomized complete block design was utilized to assign the twelve biogas reactor treatments to four blocks, within eastern and central regions of Kenya. We obtained 42 million paired-end reads that were annotated against sixteen reference databases using two ENVO ontologies, prior to β-diversities studies. We identified 37 phyla, 65 classes and 132 orders of micro-organisms. Bacteria dominated the microbiome and comprised of 28 phyla, 42 classes and 92 orders, conveying substrate’s versatility in the treatments. Though, fungi and Archaea comprised of only 5 phyla, the fungi were richer; suggesting the importance of hydrolysis and fermentation in biogas production systems. High β-diversity within the taxa was largely linked to communities’ metabolic capabilities. Clostridiales and Bacteroidales, the most prevalent guilds, metabolize organic macromolecules. The identified affiliates of Cytophagales, Alteromonadales, Flavobacteriales, Fusobacteriales, Deferribacterales, Elusimicrobiales, Chlamydiales, Synergistales to mention but few, also catabolize macromolecules into smaller substrates to conserve energy. Furthermore, δ-Proteobacteria, Gloeobacteria and Clostridia affiliates syntrophically regulate PH2 and reduce metal to provide reducing equivalents. Methanomicrobiales and other Methanomicrobia species were the most prevalence Archaea, converting formate, CO2(g), acetate and methylated substrates into CH4(g). Thermococci, Thermoplasmata and Thermoprotei were among the sulfur and other metal reducing Archaea that contributed to redox balancing and other metabolism within treatments. Eukaryotes, mainly fungi were the least abundant guild, comprised largely Ascomycota and Basidiomycota species. Chytridiomycetes, Blastocladiomycetes and Mortierellomycetes were among the rare species, suggesting their metabolic and substrates limitations. Generally, we observed that environmental and treatment perturbations influenced communities’ abundance, β-diversity and reactor performance largely through stochastic effect. The study of the diversity of the biogas’ microbiomes over wide environmental variables and the productivity of biogas reactor systems has provided insights into better management strategies that may ameliorate biochemical limitations to effective biogas production.Author SummaryThe failure of biochemical reactions in biogas producing systems is a common problem and results from poor functioning of the inhabiting micro-organisms. A poor understanding of the global diversities of these micro-organisms and lack of information on the link between environmental variables, biogas production, and community composition, contrains the development of strategies that can ameliorate these biochemical issues. We have integrated sequencing-by-synthesis technology and intensive computational approaches to reveal metacommunities in the studied reactor treatments. The identified communities were compared with the treatment’s phenotypic and environmental data in an attempt to fill the existing knowledge gaps on biogas microbiomes and their production capacities. We present 132 biogas taxonomic profiles systematically and comparatively, linking the abundance with the identified environmental variables. The local composition of microbiome and variations in abundance were also linked to the observed differences in biogas productivity, suggesting the possible cause of the observed variations. The detailed information presented in this study can aid in the genetic manipulation or formulation of optimal microbial ratios to improve their effectiveness in biogas production.

The influence of crude oil and chemical dispersant was evaluated over planktonic bacteria and biofilms grown on API 5L steel surfaces in microcosm systems. Three conditions were simulated, an untreated marine environment and a marine environment with the presence of crude oil and a containing crude oil and chemical dispersant. The results of coupon corrosion rates indicated that in the oil microcosm, there was a high corrosion rate when compared with the other two systems. Analysis of bacterial communities by 16S rRNA gene sequencing described a clear difference between the different treatments. In plankton communities, the Bacilli and Gammaproteobacteria classes were the most present in numbers of operational taxonomic unit (OTUs). The Vibrionales, Oceanospirillales, and Alteromonadales orders were predominant in the treatment with crude oil, whereas in the microcosm containing oil and chemical dispersant, mainly members of Bacillales order were detected. In the communities analyzed from biofilms attached to the coupons, the most preponderant class was Alphaproteobacteria, followed by Gammaproteobacteria. In the control microcosm, there was a prevalence of the orders Rhodobacterales, Aeromonadales, and Alteromonadales, whereas in the dispersed oil and oil systems, the members of the order Rhodobacterales were present in a larger number of OTUs. These results demonstrate how the presence of a chemical dispersant and oil influence the corrosion rate and bacterial community structures present in the water column and biofilms grown on API 5L steel surfaces in a marine environment. KEY POINTS: • Evaluation of the effects of oil and chemical surfactants on the corrosion of API 5L. • Changes in microbial communities do not present corrosive biofilm on API 5L coupons.

The Arava Valley in Israel is a rock desert within the Great African Rift valley. Soil from this area is covered with a salt crust. Here, we report microbial diversity from arid, naturally saline samples collected near Ein Yahav from the Arava Valley by culture-independent as well as culture-dependent analysis. High-throughput sequencing of the hypervariable region V4 of the 16S rRNA gene revealed that the microbial community consists of halophiles from the domain Bacteria as well as Archaea. Bacterial diversity was mainly represented by the genus Salinimicrobium of the order Flavobacteriales within the phylum Bacteroidetes, from the gammaproteobacterial orders Alteromonadales and Oceanospirillales as well as representatives from the order Bacillales of the phylum Firmicutes. Archaeal diversity was dominated by euryarchaeal Halobacteria from the orders Halobacteriales, Haloferacales and Natrialbales. But more than 40 % of the sequences affiliated with Archaea were assigned to unknown or unclassified archaea. Even if taxonomic resolution of the 16S rRNA gene V4 region for Archaea is limited, this study indicates the need of further and more detailed studies of Archaea. By using culture-dependent analysis bacteria of the order Bacillales as well as archaea from all three halobacterial orders Halobacteriales, Haloferacales and Natrialbales including potentially novel species from the genera Halorubrum and Haloparvum were isolated.

In this study, Illumina Miseq sequencing of 16S rRNA gene amplicon was performed on sediments collected from Krossfjorden, Arctic for analyzing the bacterial community structure. Metagenome contained 15,936 sequences with 5,809,491 bp size and 53% G+C content. Metagenome sequence information are now available at NCBI under the Sequence Read Archive (SRA) database with accession no. SRP159159. Taxonomic hits distribution from MG-RAST analysis revealed the dominance of Alpha- and Gamma-subdivisions of Proteobacteria (88.89%) along with Bacteriodetes (8.89%) and Firmicutes (2.22%). Predominant species were Alteromonadales bacterium TW-7 (24%), Pseudoalteromonas haloplanktis (20%) and Pseudoalteromonas spp. SM9913 (18%). MG-RAST assisted analysis also detected the presence of a variety of marine taxa like Bacteriodes, Pseudovibrio, Marinobacter, Idiomarina, Teredinibacter, etc. which take part in key ecological functions and biogeochemical activities of Arctic fjord ecosystems. Keywords: Arctic, Bacterial diversity, Metagenome, Amplicon sequencing, Illumina

A Gram-stain-negative bacterium, designated strain IO390401T, was isolated from a seawater sample from the sulphide region of the Indian Ocean. Phylogenetic trees based on 16S rRNA gene sequences showed that strain IO390401T is a member of the genus Alteromonas , sharing 97.0−97.4 % 16S rRNA gene sequence similarity with Alteromonas addita R10SW13T, A. stellipolaris LMG 21861T, A. naphthalenivorans JCM 17741T, A. gracilis 9a2T and A. australica H17T, and 94.8−96.8 % with the type strains of other species of the genus Alteromonas . Strain IO390401T contained ubiquinone-8 (Q-8) as the sole isoprenoid quinone, C16:0 and C16:1 ω7c/C16:1 ω6c as the dominant cellular fatty acids, and phosphatidylglycerol and phosphatidylethanolamine as the major polar lipids. The genome of strain IO390401T consists of a 4.4 Mb chromosome with a G+C content of 48.2 mol%. Average nucleotide identity values between strain IO390401T and the three closest phylogenetic neighbours, namely A. addita R10SW13T, A. stellipolaris LMG 21861T and A. naphthalenivorans JCM 17741T, were 70.5, 70.4 and 70.6 %, respectively, and the corresponding in silico DNA–DNA hybridization values were 20.6, 20.7 and 21.1 %. Phylogenetic distinctiveness and chemotaxonomic differences, together with phenotypic properties determined in this study, revealed that strain IO390401T could be differentiated from closely related species. It is therefore proposed as representing a novel species in the genus Alteromonas , for which the name Alteromonas indica sp. nov. is suggested. The type strain is IO390401T (=JCM 32638T=CGMCC 1.13554T=CCTCC AB 2018072T).

Analyzing microbial communities and their functional structure in an expeditious and extensive manner is very important to identify ecological indicators in the historically polluted niches. Thus, the present study aims to investigate the metabolic potential and phylogenetic diversity of oil and coal contaminated sediments of Navlkahi port (NAV), Gulf of Kachchh, Gujarat, India during two seasons viz. pre-monsoon (PreMN) and post-monsoon (PoMN) using NextSeq 499, a high throughput sequencing platform. Assessment of functional diversity of both metagenomes by SEED, Gene Ontology terms, Clusters of Orthologous Groups of protein categories, protein family numbers, and Kyoto Encyclopedia of Genes revealed xenobiotic compound degradation potential of indigenous microbial community. The results also revealed the higher abundance of genes involved in aromatic compound degradation, heavy metal resistance, stress response and other major biochemical cycles during PreMN season compared to PoMN. Taxonomic assessment suggests Alteromonadales were the predominant from phyla Proteobacteria at NAV. Apart from abundant microbial communities, “rare” community may also a play major role in the xenobiotic compound degradation. The statistical analysis indicates that the diversity and richness were almost similar during both the seasons. The existence of indicator microbial communities in xenobiotic compounds degradation and other major biochemical cycles in both contaminated metagenomes implies that stimulating indigenous microorganisms could be a valid option for remediating oil and coal contaminated sites. Moreover, the information generated from this experiment with high throughput sequencing may be the first report particularly with reference to functional and phylogenetic microbial diversity of coal and oil contaminated habitat.

Seasonal variations, as well as depth-wise distribution patterns of bacterial and archaeal communities in the Arabian Sea oxygen minimum zone (AS-OMZ), were analyzed by examining 16S rRNA gene clones and their sequences. Sampling was carried out at the Arabian Sea Time Series (ASTS) location (17°0.126′ N, 67°59.772′ E) from five different depths during three different seasons. A total of 743 and 256 non-chimeric bacterial and archaeal 16S rRNA gene sequences were analyzed. Most of the bacterial 16S rRNA gene sequences were affiliated to Gammaproteobacteria (39.31%), Alphaproteobacteria (23.56%) and Cyanobacteria (20.2%). The archaeal 16S rRNA gene sequences mostly aligned with Marine Group II (MG-II, Euryarchaeota). An explicit vertical partitioning of bacterial communities between the surface (surface and DCM) and OMZ (250 m, 500 m, and 1000 m) was observed. Also evident was an apparent seasonal variation among surface bacterial communities but a minimal variation among OMZ bacterial and archaeal communities. LINKTREE and canonical correspondence analysis (CCA) indicates that differences in the concentrations of dissolved oxygen (DO) and total organic carbon (TOC) seem to cause the vertical separation among bacterial communities in the ASTS station. A higher number of shared OTUs affiliated mainly to Alteromonadales (bacteria) and Methanosarcinales (archaea) contributed towards seasonally stable community structure in the OMZ depths.

The abundance and composition of bacterioplankton of the Northern South China Sea (NSCS) were investigated using flow cytometry and high-throughput sequencing. The results showed that the absolute abundance of bacterioplankton retained high values in surface waters at both continental shelf and oceanic sites and Proteobacteria, Cyanobacteria, and Bacteroidetes represented the three typical dominant phyla in NSCS. The average bacterioplankton abundances at 5 m, 75 m, and 200 m were 9.55, 5.04, and 1.32 × 105 cells mL−1, respectively, and there was a significantly positive correlation between bacterioplankton abundance and Chl a content (r = 0.84, p

A bacterial strain designated LSN-49T was isolated from a brackish river in Taiwan and characterized using a polyphasic taxonomy approach. Cells of strain LSN-49T were Gram-staining-negative, aerobic, poly-β-hydroxybutyrate accumulating, motile by means of a monopolar flagellum, non-spore forming, straight rods and formed shiny and translucent colonies. Growth occurred at 20–40 °C (optimum, 25–30 °C), at pH 6–10 (optimum, pH 7–8) and with 0–3 % (w/v) NaCl [optimum, 0–1 % (w/v)]. The predominant fatty acids were summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c), C17 : 1ω8c and C16 : 0. The polar lipid profile consisted of a mixture of phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylcholine, (PC), two uncharacterized aminophospholipids (APL1 and APL2), one uncharacterized glycolipid (GL1), four uncharacterized phospholipids (PL1–PL4) and four uncharacterized lipids (L1–L4). The major polyamine was putrescine. The major isoprenoid quinone was Q-8 and the DNA G+C content was 51.0 mol%. The results of phylogenetic analyses based on 16S rRNA gene sequences indicated that LSN-49T formed a distinct lineage with respect to closely related genera in the family Pseudoalteromonadaceae . LSN-49T was most closely related to Pseudoalteromonas , Algicola and Psychrosphaera and showed 89.3–92.1 % sequence similarity with members of the family Pseudoalteromonadaceae with validly published names. On the basis of the genotypic and phenotypic data, LSN-49T represents a novel genus and species of the family Pseudoalteromonadaceae , for which the name Salsuginimonas clara gen. nov., sp. nov. is proposed. The type strain is LSN-49T (=BCRC 81005T=LMG 29726T=KCTC 52439T).

The Meloidogyne-based disease complexes (MDCs) are caused by the interaction of different root-knot nematode species and phytopathogenic fungi. These complexes are devastating several important crops worldwide including tomato and coffee. Despite their relevance, little is known about the role of the bacterial communities in the MDCs. In this study 16s rDNA gene sequencing was used to analyze the bacterial microbiome associated with healthy and infested roots, as well with females and eggs of Meloidogyne enterolobii and M. paranaensis, the causal agents of MDC in tomato and coffee, respectively. Each MDC pathosystems displayed a specific taxonomic diversity and relative abundances constituting a very complex system. The main bacterial drivers of the MDC infection process were identified for both crops at order level. While corky-root coffee samples presented an enrichment of Bacillales and Burkholderiales, the corcky-root tomato samples presented an enrichment on Saprospirales, Chthoniobacterales, Alteromonadales, and Xanthomonadales. At genus level, Nocardia was common to both systems, and it could be related to the development of tumor symptoms by altering both nematode and plant systems. Furthermore, we predicted the healthy metabolic profile of the roots microbiome and a shift that may result in an increment of activity of central metabolism and the presence of pathogenic genes in both crops., The research leading to these results has received funding from the Research Projects of High Strategic Value for Society (Project 2003530920) founded by Institute of Ecology A. C (INECOL).

The phytoplanktonic production and prokaryotic consumption of organic matter significantly contribute to marine carbon cycling. Organic matter released from phytoplankton via three processes (exudation of living cells, cell disruption through grazing, and viral lysis) shows distinct chemical properties. We herein investigated the effects of phytoplanktonic whole-cell fractions (WF) (representing cell disruption by grazing) and extracellular fractions (EF) (representing exudates) prepared from Heterosigma akashiwo, a bloom-forming Raphidophyceae, on prokaryotic communities using culture-based experiments. We analyzed prokaryotic community changes for two weeks. The shift in cell abundance by both treatments showed similar dynamics, reaching the first peak (~4.1×106‍ ‍cells‍ ‍mL–1) on day 3 and second peak (~1.1×106‍ ‍cells‍ ‍mL–1) on day 13. We classified the sequences obtained into operational taxonomic units (OTUs). A Bray-Curtis dissimilarity analysis revealed that the OTU-level community structure changed distinctively with the two treatments. Ten and 13 OTUs were specifically abundant in the WF and EF treatments, respectively. These OTUs were assigned as heterotrophic bacteria mainly belonging to the Alteromonadales (Gammaproteobacteria) and Bacteroidetes clades and showed successive dynamics following the addition of organic matter. We also analyzed the dynamics of these OTUs in the ocean using publicly available metagenomic data from a natural coastal bloom in Monterey Bay, USA. At least two WF treatment OTUs showed co-occurrence with H. akashiwo, indicating that the blooms of H. akashiwo also affect these OTUs in the ocean. The present results strongly suggest that the thriving and dead cells of uninfected phytoplankton differentially influence the marine prokaryotic community.

In the present research, bacterial diversity was studied during a 6-month feeding trial utilizing zebrafish (Danio rerio) fed Hermetia illucens reared on different substrates with an emphasis on fish gut bacterial diversity. A polyphasic approach based on viable counting, PCR-DGGE and metagenomic 16S rRNA gene amplicon target sequencing was applied. Two different H. illucens groups were reared on coffee by-products (C) or a mixture of vegetables (S). Viable counts showed a wide variability based on substrate. PCR-DGGE and Illumina sequencing allowed the major and minor bacterial taxa to be detected. Both samples of larvae and their frass reared on the S substrate showed the highest richness and evenness of bacterial communities, whereas zebrafish (ZHC) fed H. illucens reared on substrate C and zebrafish (ZHS) fed H. illucens reared on substrate S had the lowest bacterial richness and evenness. A stimulating effect of bioactive compounds from coffee by-products on the occurrence of Lactobacillaceae and Leuconostoccaceae in H. illucens reared on substrate C has been hypothesized. Zebrafish gut samples originating from the two feeding trials showed complex microbial patterns in which Actinobacteria and Alteromonadales were always detected, irrespective of the diet used. Enterobacteriaceae in fish guts were more abundant in ZHS than in ZHC, thus suggesting an influence of the bioactive compounds (chlorogenic and caffeic acids) in the substrate on Enterobacteriaceae in fish guts. ZHC showed a higher abundance of Clostridia than did ZHS, which was likely explained by stimulating activity on the bacteria in this class by the bioactive compounds contained in H. illucens reared on substrate C. An influence of the microbiota of H. illucens or insect-derived bioactive compounds on the gut microbiota of zebrafish has been suggested. The presence of bacteria consistently associated with zebrafish guts has been found irrespective of the diet, thus attesting to the likely stability of the core fish microbiota.

Diversity and abundance of the denitrifying genes nirK, nirS and nosZ were investigated in cow manure compost using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time quantitative PCR (qPCR), respectively. These three genes were detected in all the stages of the composting process. Phylogenetic analysis showed that the nirK gene was closely related to Rhizobiales, Burkholderiales, the nirS gene was closely related to Pseudomonadales and Burkholderiales, and the nosZ gene was closely related to Rhodospirillales, Rhizobiales, Pseudomonadales, and Alteromonadales. qPCR results showed that the abundance of these three genes (nirK, nirS and nosZ) reached the peak value in the late thermophilic stage of composting and abundance of the nirK gene was higher than that of the nosZ gene and the nirS gene. Redundancy analysis (RDA) showed that the diversity of the nirK and nirS genes was significantly correlated with ammonium (p < 0.05), the diversity of the nosZ gene was significantly correlated with pH (p < 0.05) and the abundance of the nirK nirS and nosZ genes was significantly correlated with temperature (p< 0.05). La diversidad y la abundancia de los genes desnitrificadores nirK, nirS, nosZ en el compost de estiércol de vaca se investigaron por medio de la reacción en cadena de la polimerasa seguida de electroforesis en gel con gradiente de desnaturalización (PCR-DGGE) y por PCR cuantitativa (qPCR) en tiempo real, respectivamente. Estos 3 genes fueron detectados durante todas las fases del compostaje. El análisis filogenético mostró estrecha relación del gen nirK con Rhizobiales y Burkholderiales, del gen nirS con Pseudomonadales y Burkholderiales y del gen nosZ con Rhodospirillales, Rhizobiales, Pseudomonadales y Alteromonadales. Los resultados de la qPCR mostraron que la abundancia de los genes nirK, nirSy nosZ alcanzó el valor máximo en la fase termofÃlica tardÃa del compostaje, y que la abundancia del gen nirK era más elevada que los de los genes nosZ y nirS. El análisis de redundancia (RDA) mostró que la diversidad de los genes nirK y nirS estaba significativamente correlacionada con la concentración de amonio (p

Summary Phytoplankton-derived polysaccharide microgels, including transparent exopolymer particles (TEP), are a major component of the marine organic carbon pool. Previous studies have made correlative links between phytoplankton material and bacterioplankton, and performed experiments that assess general responses to phytoplankton, yet there is a lack of direct empirical evidence of specific bacterioplankton responses to natural phytoplankton polysaccharide microgels. In this study, we used diatom produced TEP in controlled incubation experiments to determine the impact of polysaccharide microgels on a coastal bacterioplankton community. Quantification of bacterial 16S rRNA gene transcripts showed that the addition of TEP caused an increase in bacterioplankton activity. Similarly, high-throughput sequencing of RT-PCR amplified bacterial 16S rRNA gene transcripts showed that active bacterioplankton community structure and diversity also changed in response to microgels. Alteromonadales and Rhodobacterales increased in abundance in response to TEP, suggesting that both bacterioplankton taxa utilize diatom-derived microgels. However, through assessing 13C-labelled TEP uptake via RNA Stable Isotope Probing, we show that only the Alteromonadales (genus Alteromonas) assimilated the TEP carbon. This study adds utilization of diatom-derived TEP to the metabolic repertoire of the archetypal copiotrophic bacterioplankton Alteromonas, and indicates that the Rhodobacterales may utilize TEP for other purposes (e.g. attachment sites).

A Gram-stain-negative, aerobic, motile and rod-shaped bacterial strain, designated RA2-7T, was isolated from a mussel (Mytilus edulis) collected from the South Sea, South Korea, and subjected to a taxonomic study using a polyphasic approach. Strain RA2-7T grew optimally at 20 °C, at pH 7.0–8.0 and in the presence of 2.0–3.0 % (w/v) NaCl. The phylogenetic trees based on 16S rRNA gene sequences showed that strain RA2-7T belonged to the genus Colwellia . Strain RA2-7T exhibited 16S rRNA gene sequence similarity values of 98.3, 98.0 and 97.5 % to the type strains of Colwellia sediminilitoris , Colwellia aestuarii and Colwellia polaris , respectively, and of 94.5–96.5 % to the type strains of the other species of the genus Colwellia . Strain RA2-7T contained Q-8 as the predominant ubiquinone and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and C16 : 0 as the major fatty acids. The major polar lipids detected in strain RA2-7T were phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content of strain RA2-7T was 39.0±0.04 mol% and its DNA–DNA relatedness values with the type strains of C . sediminilitoris , C. aestuarii and C. polaris were 14–19 %. Differential phenotypic properties, together with its phylogenetic and genetic distinctiveness, revealed that strain RA2-7T is separated from recognized species of the genus Colwellia . On the basis of the data presented, strain RA2-7T is considered to represent a novel species of the genus Colwellia , for which the name Colwellia mytili sp. nov. is proposed. The type strain is RA2-7T (=KCTC 52417T=NBRC 112381T).

The blue mussel genus Mytilus is a popular food source and an important model organism for many scientific disciplines [1,2]. The morphologically similar species M. edulis (Me) and M. trossulus (Mt) form an unprecedented and unique hybridisation pattern in the Baltic Sea [3, 4]. Recently, it was shown that metabolite profiling of Mytilus species provides significant potential for biomonitoring of aquatic environments but also for informing natural product research [1,2]. However, the metabolome of Mytilus sp. is still little investigated and no metabolome data from the Baltic Sea blue mussels are available so far. In order to compare the metabolite profiles of Baltic Mytilus sp. at an individual level, 30 Mytilus specimens were sampled at a blue mussel farm in Kiel (Baltic Sea) and genotyped with two commonly utilized nuclear markers, EFbis and Glu-5' [4]. Genotypic assessment revealed a dominance of Mt alleles at Kiel Fjord with 90% of the samples being intermediate (Me/Mt) or Mt-like hybrids. Secondary metabolites of the whole tissue extracts (EtOAc) were explored by HPLC-DAD-MS. Metabolite profiles were found to be similar but not identical, differing mainly in minor nonpolar secondary metabolites. Considering that secondary metabolites are often produced by associated microbes and not by the macroorganism itself, the microbiome of the mussel specimens was also investigated. A culture-based approach yielded 259 bacterial and fungal strains, including Actinomycetes. Bacteria belonging to the orders Alteromonadales and Pseudomonadales (Acinetobacter sp., Pseudomonas sp., Pseudoalteromonas sp., Psychrobacter sp., Shewanella sp.) dominated the microbial community (57%). In addition, an RFLP based culture-independent approach was developed for Mytilus sp. individuals. Both the culture-dependent and culture-independent approaches showed inter-individual differences with respect to the associated microbes, but no genotypic dependence.

Infectious diseases such as white plague syndrome (WPS) and black band disease (BBD) have caused massive coral loss worldwide. We performed a metaproteomic study on the Abrolhos coral Mussismilia braziliensis to define the types of proteins expressed in healthy corals compared to WPS- and BBD-affected corals. A total of 6363 MS/MS spectra were identified as 361 different proteins. Healthy corals had a set of proteins that may be considered markers of holobiont homoeostasis, including tubulin, histone, Rab family, ribosomal, peridinin-chlorophyll a-binding protein, F0F1-type ATP synthase, alpha-iG protein, calmodulin and ADP-ribosylation factor. Cnidaria proteins found in healthy M. braziliensis were associated with Cnidaria-Symbiodinium endosymbiosis and included chaperones (hsp70, hsp90 and calreticulin), structural and membrane modelling proteins (actin) and proteins with functions related to intracellular vesicular traffic (Rab7 and ADP-ribosylation factor 1) and signal transduction (14-3-3 protein and calmodulin). WPS resulted in a clear shift in the predominance of proteins, from those related to aerobic nitrogen-fixing bacteria (i.e. Rhizobiales, Sphingomonadales and Actinomycetales) in healthy corals to those produced by facultative/anaerobic sulphate-reducing bacteria (i.e. Enterobacteriales, Alteromonadales, Clostridiales and Bacteroidetes) in WPS corals. BBD corals developed a diverse community dominated by cyanobacteria and sulphur cycle bacteria. Hsp60, hsp90 and adenosylhomocysteinase proteins were produced mainly by cyanobacteria in BBD corals, which is consistent with elevated oxidative stress in hydrogen sulphide- and cyanotoxin-rich environments. This study demonstrates the usefulness of metaproteomics for gaining better comprehension of coral metabolic status in health and disease, especially in reef systems such as the Abrolhos that are suffering from the increase in global and local threatening events.

Deep-sea surface sediments and flocculent material (floc) associated with corals containing oil originating from the Deepwater Horizon (DWH) oil spill were examined to determine the diversity of microbes and the presence of functional genes involved in oil degradation. For all samples, 16S rRNA clone libraries were constructed to obtain full-length sequences and Illumina amplicon sequencing was used to further probe the diversity of the microbial community. The 16S rRNA gene data obtained by Illumina amplicon sequencing revealed Proteobacteria (55–64%) as the dominant bacteria in both sediment and floc samples. The floc samples were comprised of mostly aerobic or facultative aerobic phylotypes including Rhizobiales, Rhodobacterales, Sphingomonadales, Rickettsiales, Alteromonadales, Pseudomonadales, whereas mixtures of the aforementioned aerobic species and anaerobic phylotypes such as Desulfobacterales, Desulfuromonadales and Desulfarculales were present in the sediment samples. Genera affiliated with oil-degrading bacteria were identified in both sediment and floc samples. To evaluate the potential of the microbial community to degrade oil, clone libraries were constructed for the alkB gene (one of the structural genes of alkane hydroxylase involved in the aerobic degradation of n-alkanes of chain length >C5–C16) and the alkylsuccinate synthase/benzylsuccinate synthases (assA/bssA) gene (involved in the anaerobic degradation of n-alkanes [via assA] and polycyclic aromatic hydrocarbons [PAHs; via bssA]). The alkB gene was present in all samples with the majority of sequences clustering to members of the Proteobacteria closely aligned to environmental sequences from hydrocarbon seep environments. The assA/bssA genes were only detected in sediment samples and were closely affiliated with δ-Proteobacteria previously detected in oil-contaminated sediments and oil-enrichment cultures. These data provide insight into the differences between environments impacted by the DWH oil spill and highlight the functional diversity of oil-degrading microbes associated with a deep-sea coral community.

Ulvan is the main polysaccharide component of the Ulvales (green seaweed) cell wall. It is composed of disaccharide building blocks comprising 3-sulfated rhamnose linked to d-glucuronic acid (GlcUA), l-iduronic acid (IdoUA), or d-xylose (Xyl). The degradation of ulvan requires ulvan lyase, which catalyzes the endolytic cleavage of the glycoside bond between 3-sulfated rhamnose and uronic acid according to a β-elimination mechanism. The first characterized ulvan lyase was identified in Nonlabens ulvanivorans, an ulvanolytic bacterial isolate. In the current study, we have identified and biochemically characterized novel ulvan lyases from three Alteromonadales isolated bacteria. Two homologous ulvan lyases (long and short) were found in each of the bacterial genomes. The protein sequences have no homology to the previously reported ulvan lyases and therefore are the first representatives of a new family of polysaccharide lyases. The enzymes were heterologously expressed in Escherichia coli to determine their mode of action. The heterologous expressed enzymes were secreted into the milieu subsequent to their signal sequence cleavage. An endolytic mode of action was observed and studied using gel permeation chromatography and (1)H NMR. In contrast to N. ulvanivorans ulvan lyase, cleavage occurred specifically at the GlcUA residues. In light of the genomic context and modular structure of the ulvan lyase families identified to date, we propose that two ulvan degradation pathways evolved independently.