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Marine nitrogen (N SB 2 sb ) fixation is important in the global biogeochemical cycling of N, and the interlinked cycling of carbon (C). 1, 2 show that qPCR data can be related to cell numbers and certainly are not consistent with the wide range of conversion factors for I nifH i gene copy: cell used by Meiler et al. ([28]). This approach generated ranges for depth integrated I nifH i gene abundance and I nifH i inferred cell counts and biomass across oceanic regions and latitudinal gradients for comparison with biogeochemical model output. [Extracted from the article]

Abstract Microbial communities are the main drivers of biogeochemical cycling of multiple elements sustaining life in the ocean. The rapidity of their response to stressors and abrupt environmental changes implies that even fast and infrequent events can affect local transformations of organic matter and nutrients. Modern molecular techniques now allow for monitoring of microbial activities and functions in the environment through the analysis of genes and expressed genes contained in natural microbial assemblages. However, messenger RNA turnover in cells can be as short as 30 seconds and stability varies greatly between transcripts. Sampling of in situ communities involves an inevitable delay between the collection of seawater and the extraction of its RNA, leaving the bacterial communities plenty of time to alter their gene expression. The characteristics of microbial RNA turnover make time-series very difficult because samples need to be processed immediately to limit alterations to the metatranscriptomes. To address these challenges we designed an autonomous in situ fixation multi-sampler (AFISsys) for the reliable sampling of microbial metatranscriptomes at frequent intervals, for refined temporal resolution. To advance the development of this instrument, we examined the minimal seawater volume necessary for adequate coverage of community gene expression, and the suitability of phenol/ethanol fixation for immediate and long-term preservation of transcripts from a microbial community. We then evaluated the field eligibility of the instrument itself, with two case studies in a brackish system. AFISsys is able to collect, fix, and store water samples independently at a predefined temporal resolution. Phenol/ethanol fixation can conserve metatranscriptomes directly in the environment for up to a week, for later analysis in the laboratory. Thus, the AFISsys constitutes an invaluable tool for the integration of molecular functional analyses in environmental monitoring in brackish waters and in aquatic environments in general. Graphical abstract Image 1 Highlights • An autonomous water in situ fixation multi-sampler (AFISsys) has been designed. • AFISsys collects, fixes, stores water samples independently off-shore at high temporal resolution. • AFISsys conserves sensitive mRNA directly in the environment for up to a week. • Metatranscriptome analysis feasible with 250 mL of water volumes. • Integration of molecular functional analyses in environmental monitoring in aquatic environments feasible. [ABSTRACT FROM AUTHOR]

Specific regions of otherwise oligotrophic oceans seem to attract fish spawning and sustain significant abundances of fish larvae. The Sargasso Sea in the North Atlantic subtropical gyre is known as the spawning area of the Atlantic eels, but numerous other fish species also spawn in the area. In order to evaluate spatial variability of larval fish in the region, we examined species diversity, composition and abundances at eight stations in the Subtropical Convergence Zone ( STCZ) using morphological identification and DNA barcoding. From a total of approximately 3500 specimens collected, at least 154 species from 50 families could be identified. The family Myctophidae had the highest species richness, with at least 32 species represented. The myctophids Lepidophanes gaussi, Bolinichthys indicus, Notolychnus valdiviae and Ceratoscopelus warmingii were the four most abundant species. Other common species included the three eels: Nemichthys scolopaceus, Ariosoma balearicum and Anguilla anguilla. Larval fish species composition differed substantially between the relatively closely spaced stations on either side of prominent hydrographic fronts in the study area, presumably because of the strong environmental gradients. Common eel species were concentrated between the fronts whereas common myctophids were of highest abundance at the outer edges of the fronts. The abundances of most species were generally enhanced in the vicinity of the fronts. The use of combined morphological and DNA-barcoding identification methods facilitated species identification, and we could document substantially higher levels and a larger degree of spatial variability in species diversity of fish larvae than previously shown for oligotrophic ocean areas. [ABSTRACT FROM AUTHOR]

Despite the paramount importance of bacteria for biogeochemical cycling of carbon and nutrients, little is known about the potential effects of climate change on these key organisms. The consequences of the projected climate change on bacterioplankton community dynamics were investigated in a Baltic Sea spring phytoplankton bloom mesocosm experiment by increasing temperature with 3°C and decreasing pH by approximately 0.4 units via CO2 addition in a factorial design. Temperature was the major driver of differences in community composition during the experiment, as shown by denaturing gradient gel electrophoresis ( DGGE) of amplified 16 S rRNA gene fragments. Several bacterial phylotypes belonging to Betaproteobacteria were predominant at 3° C but were replaced by members of the Bacteriodetes in the 6°C mesocosms . Acidification alone had a limited impact on phylogenetic composition, but when combined with increased temperature, resulted in the proliferation of specific microbial phylotypes. Our results suggest that although temperature is an important driver in structuring bacterioplankton composition, evaluation of the combined effects of temperature and acidification is necessary to fully understand consequences of climate change for marine bacterioplankton, their implications for future spring bloom dynamics, and their role in ecosystem functioning. [ABSTRACT FROM AUTHOR]

Variation in traits causes bacterial populations to respond in contrasting ways to environmental drivers. Learning about this will help us understand the ecology of individual populations in complex ecosystems. We used 454 pyrosequencing of the hypervariable region V6 of the 16S rRNA gene to study seasonal dynamics in Baltic Sea bacterioplankton communities, and link community and population changes to biological and chemical factors. Surface samples were collected from May to October 2003 and in May 2004 at the Landsort Deep in the central Baltic Sea Proper. The analysis rendered, on average, 20 200 sequence reads for each of the eight samples analyzed, providing the first detailed description of Baltic Sea bacterial communities. Community composition varied dramatically over time, supporting the idea of strong temporal shifts in bacterioplankton assemblages, and clustered according to season (including two May samples from consecutive years), suggesting repeatable seasonal succession. Overall, community change was most highly correlated with change in phosphorus concentration and temperature. Individual bacterial populations were also identified that tightly co-varied with different Cyanobacteria populations. Comparing the abundance profiles of operational taxonomic units at different phylogenetic distances revealed a weak but significant negative correlation between abundance profile similarity and genetic distance, potentially reflecting habitat filtering of evolutionarily conserved functional traits in the studied bacterioplankton. [ABSTRACT FROM AUTHOR]

Through lysis of bacterioplankton cells, viruses mediate an important, but poorly understood, pathway of carbon and nutrients from the particulate to the dissolved form. Via this activity, nutrient-rich cell lysates may become available to noninfected cells and support significant growth. However, the nutritional value of lysates for noninfected bacteria presumably depends on the prevailing nutrient limitation. In the present study, we examined dynamics of dissolved DNA (D-DNA) and viruses along a transect in the phosphorus (P)-limited Öre Estuary, northern Baltic Sea. We found that viruses were an important mortality factor for bacterioplankton and that their activity mediated a significant recycling of carbon and especially of P. Uptake of dissolved DNA accounted for up to 70% of the bacterioplankton P demand, and about a quarter of the D-DNA pool was supplied through viral lysis of bacterial cells. Generally, the importance of viral lysates and uptake of D-DNA was highest at the estuarine and offshore stations and was positively correlated with P limitation measured as alkaline phosphatase activity. Our results highlight the importance of viral activity for the internal recycling of principal nutrients and pinpoints D-DNA as a particularly relevant compound in microbial P dynamics. [ABSTRACT FROM AUTHOR]

Bacteria growing on marine particles generally have higher densities and cell-specific activities than free-living bacteria. Since rapidity of phage adsorption is dependent on host density, while infection productivity is a function of host physiological status, we hypothesized that marine particles are sites of elevated phage production. In the present study, organic-matter-rich agarose beads and a marine phage–host pair ( Cellulophaga sp., ΦSM) were used as a model system to examine whether bacterial colonization of particles increases phage production. While no production of phages was observed in plain seawater, the presence of beads enhanced attachment and growth of bacteria, as well as phage production. This was observed because of extensive lysis of bacteria in the presence of beads and a subsequent increase in phage abundance both on beads and in the surrounding water. After 12 h, extensive phage lysis reduced the density of attached bacteria; however, after 32 h, bacterial abundance increased again. Reexposure to phages and analyses of bacterial isolates suggested that this regrowth on particles was by phage-resistant clones. The present demonstration of elevated lytic phage production associated with model particles illustrates not only that a marine phage has the ability to successfully infect and lyse surface-attached bacteria but also that acquisition of resistance may affect temporal phage–host dynamics on particles. These findings from a model system may have relevance to the distribution of phage production in environments rich in particulate matter (e.g., in coastal areas or during phytoplankton blooms) where a significant part of phage production may be directly linked to these nutrient-rich “hot spots.” [ABSTRACT FROM AUTHOR]

Cyanobacteria are regarded as the main N2-fixing organisms in marine waters. However, recent clone libraries from various oceans show a wide distribution of the dinitrogenase reductase gene ( nifH) originating from heterotrophic bacterioplankton. We isolated heterotrophic N2-fixing bacteria from Baltic Sea bacterioplankton using low-nitrogen plates and semi-solid diazotroph medium (SSDM) tubes. Isolates were analysed for the nitrogenase ( nifH) gene and active N2 fixation by nested polymerase chain reaction (PCR) and acetylene reduction respectively. A primer-probe set targeting the nifH gene from a γ -proteobacterial isolate, 97% 16S rDNA similarity to Pseudomonas stutzeri, was designed for measuring in situ dynamics using quantitative real-time PCR. This nifH gene sequence was detected at two of 11 stations in a Baltic Proper transect at abundances of 3 × 104 and 0.8 × 103 copies per litre seawater respectively. Oxygen requirements of isolates were examined by cultivation in SSDM tubes where oxygen gradients were determined with microelectrodes. Growth, and thereby N2 fixation, was observed as horizontal bands formed at oxygen levels of 0–6% air saturation. The apparent microaerophilic or facultative anaerobic nature of the isolates explains why the SSDM approach is the most appropriate isolation method. Our study illustrates how combined isolation, functional analyses and in situ quantification yielded insights into the oxygen requirements of heterotrophic N2-fixing bacterioplankton isolates, which were confirmed to be present in situ. [ABSTRACT FROM AUTHOR]

Studies the widespread N-acetyl-D-glucosamine (NAG) uptake among pelagic marine bacteria and its ecological implications. NAG uptake in isolates; Streptozotocin sensitivity in isolates; Facultative anaerobics among isolates.

Examines bacterial community composition, enzymatic activities and carbon dynamics during diatom blooms in four laboratory seawater mesocosms. Effect of colonization and hydrolysis of decaying phytoplankton on bacterial community composition; Chlorophyll a and microbial abundances; Bacterial production and specific growth rates.

Provides information on a study which examined horizontal and vertical variations in bacterial community compositions in samples collected during the two Joint Global Ocean Flux Study Arabian Sea cruises during the Northeast monsoon in 1995. Methods; Results and discussion.

Seagrass meadows play pivotal roles in coastal biochemical cycles, with nitrogen fixation being a well-established process associated with living seagrass. Here, we tested the hypothesis that nitrogen fixation is also associated with seagrass debris in Danish coastal waters. We conducted a 52-day in situ experiment to investigate nitrogen fixation (proxied by acetylene reduction) and dynamics of the microbial community (16S rRNA gene amplicon sequencing) and the nitrogen fixing community (nifH DNA/RNA amplicon sequencing) associated with decomposing Zostera marina leaves. The leaves harboured distinct microbial communities, including distinct nitrogen fixers, relative to the surrounding seawater and sediment throughout the experiment. Nitrogen fixation rates were measurable on most days, but highest on days 3 (dark, 334.8 nmol N g−1 dw h−1) and 15 (light, 194.6 nmol N g−1 dw h−1). Nitrogen fixation rates were not correlated with the concentration of inorganic nutrients in the surrounding seawater or with carbon:nitrogen ratios in the leaves. The composition of nitrogen fixers shifted from cyanobacterial Sphaerospermopsis to heterotrophic genera like Desulfopila over the decomposition period. On the days with highest fixation, nifH RNA gene transcripts were mainly accounted for by cyanobacteria, in particular by Sphaerospermopsis and an unknown taxon (order Nostocales), alongside Proteobacteria. Our study shows that seagrass debris in temperate coastal waters harbours substantial nitrogen fixation carried out by cyanobacteria and heterotrophic bacteria that are distinct relative to the surrounding seawater and sediments. This suggests that seagrass debris constitutes a selective environment where degradation is affected by the import of nitrogen via nitrogen fixation. [ABSTRACT FROM AUTHOR]

Diazotrophs provide a significant reactive nitrogen source in the ocean. Increased warming and stratification may decrease nutrient availability in the future, forcing microbial communities toward using dissolved organic matter (DOM). Not depending on reactive nitrogen availability, diazotrophs may be "winners" in a nutrient‐depleted ocean. However, their ability to exploit DOM may influence this success. We exposed cultures of the widespread Crocosphaera to low (26°C, pH 8.1), moderate (28°C, pH 8.0), and extreme (30°C, pH 7.9) climate change scenarios, under control or DOM‐amended conditions. Growth was suboptimal in the low and extreme treatments and favored in the moderate treatment. DOM was preferred as a carbon source regardless of the treatment and promoted N2 fixation in extreme conditions. This was reflected in the increased expression of photosynthesis genes to obtain energy. DOM provides Crocosphaera with a key ecological advantage, possibly dictating diazotroph‐derived nitrogen inputs in the future ocean. [ABSTRACT FROM AUTHOR]

The oligotrophic subtropical gyre covers a vast area of the Atlantic Ocean. Decades of time-series monitoring have generated detailed temporal information about zooplankton species and abundances at fixed locations within the gyre, but their live/dead status is often omitted, especially in the dynamic subtropical convergence zone (STCZ) where the water column stratification pattern can change considerably across the front as warm and cold water masses converge. We conducted a detailed survey in the North Atlantic STCZ and showed that over 85% of the copepods were typically concentrated in the upper 200 m. Copepod carcasses were present in all samples and their proportional numerical abundances increased with depth, reaching up to 91% at 300–400 m. Overall, 14–19% of the copepods within the upper 200 m were carcasses. Shipboard experiments showed that during carcass decomposition, microbial respiration increased, and the bacterial community associated with the carcasses diverged from that in the ambient water. Combining field and experimental data, we estimated that decomposing copepod carcasses constitute a negligible oxygen sink in the STCZ, but sinking carcasses may represent an overlooked portion of the passive carbon sinking flux and should be incorporated in future studies of carbon flux in this area. [ABSTRACT FROM AUTHOR]

Non-cyanobacterial diazotrophs (NCDs) are widespread and active in marine waters. The carbon and low-oxygen (O2) conditions required for their N2 fixation may be encountered on marine particles, while a putative role of light remains uninvestigated. This study explored factors that regulate N2 fixation in Rhodopseudomonas sp. BAL398--a anoxygenic phototrophic bacterium isolated from low-salinity surface waters. Light (250 µmol photons m-2 s-1) and anoxia (0 µM O2) stimulated growth and N2 fixation; however, diazotrophy in light was dependent on high organic carbon levels (35 mM, glucose:succinate). Immunolabeling revealed that cellular nitrogenase levels increased with light, decreasing inorganic nitrogen (N) and ambient O2 (250 µM). Light and O2 stimulated motility and biofilm formation on surfaces, and N2 fixation rates increased compared to the control treatment. N2 fixation rates were positively correlated with the formation of rosette-like cellular structures, and an increased concentration of nitrogenase was observed toward the center of these structures, which increased their occurrence 600 times when cultures reached maximum N2 fixation rates vs when they had low rates. Interestingly, N2 fixation was not completely inhibited under oxic conditions and was accompanied by increased formation of capsules and cysts. Rosettes, as well as capsules and cysts, may thus serve as protection against O2. Our study reveals the physiological adaptations that underlie N2 fixation in an anoxygenic phototroph, emphasizing the significance of biofilm formation for utilizing light and fixing N2 under oxic conditions, and underscores the need for deciphering the importance of light for marine NCDs. IMPORTANCE The contribution of non-cyanobacterial diazotrophs (NCDs) to total N2 fixation in the marine water column is unknown, but their importance is likely constrained by the limited availability of dissolved organic matter and low O2 conditions. Light could support N2 fixation and growth by NCDs, yet no examples from bacterioplankton exist. In this study, we show that the phototrophic NCD, Rhodopseudomonas sp. BAL398, which is a member of the diazotrophic community in the surface waters of the Baltic Sea, can utilize light. Our study highlights the significance of biofilm formation for utilizing light and fixing N2 under oxic conditions and the role of cell plasticity in regulating these processes. Our findings have implications for the general understanding of the ecology and importance of NCDs in marine waters. [ABSTRACT FROM AUTHOR]

Nitrogen fixation provides bioavailable nitrogen, supporting global ecosystems and influencing global cycles of other elements. It provides an additional source of nitrogen to organisms at a cost of lower growth efficiency, largely due to respiratory control of intra-cellular oxygen. Nitrogen-fixing bacteria can, however, utilize both dinitrogen gas and fixed nitrogen, decreasing energetic costs. Here we present an idealized metabolic model of the heterotrophic nitrogen fixer Azotobacter vinelandii which, constrained by laboratory data, provides quantitative predictions for conditions under which the organism uses either ammonium or nitrogen fixation, or both, as a function of the relative supply rates of carbohydrate, fixed nitrogen as well as the ambient oxygen concentration. The model reveals that the organism respires carbohydrate in excess of energetic requirements even when nitrogen fixation is inhibited and respiratory protection is not essential. The use of multiple nitrogen source expands the potential niche and range for nitrogen fixation. The model provides a quantitative framework which can be employed in ecosystem and biogeochemistry models. [ABSTRACT FROM AUTHOR]

The Arctic Ocean is particularly affected by climate change with unknown consequences for primary productivity. Diazotrophs—prokaryotes capable of converting atmospheric nitrogen to ammonia—have been detected in the often nitrogen-limited Arctic Ocean but distribution and community composition dynamics are largely unknown. We performed amplicon sequencing of the diazotroph marker gene nifH from glacial rivers, coastal, and open ocean regions and identified regionally distinct Arctic communities. Proteobacterial diazotrophs dominated all seasons, epi- to mesopelagic depths and rivers to open waters and, surprisingly, Cyanobacteria were only sporadically identified in coastal and freshwaters. The upstream environment of glacial rivers influenced diazotroph diversity, and in marine samples putative anaerobic sulphate-reducers showed seasonal succession with highest prevalence in summer to polar night. Betaproteobacteria (Burkholderiales, Nitrosomonadales, and Rhodocyclales) were typically found in rivers and freshwater-influenced waters, and Delta- (Desulfuromonadales, Desulfobacterales, and Desulfovibrionales) and Gammaproteobacteria in marine waters. The identified community composition dynamics, likely driven by runoff, inorganic nutrients, particulate organic carbon, and seasonality, imply diazotrophy a phenotype of ecological relevance with expected responsiveness to ongoing climate change. Our study largely expands baseline knowledge of Arctic diazotrophs—a prerequisite to understand underpinning of nitrogen fixation—and supports nitrogen fixation as a contributor of new nitrogen in the rapidly changing Arctic Ocean. [ABSTRACT FROM AUTHOR]

The nitrogen input through biological N 2 fixation is essential for life in vast areas of the global ocean. The belief is that cyanobacteria are the only relevant N 2 -fixing (diazotrophic) organisms. It has, however, now become evident that non-cyanobacterial diazotrophs, bacteria and archaea with ecologies fundamentally distinct from those of cyanobacteria, are widespread and occasionally fix N 2 at significant rates. The documentation of a globally relevant nitrogen input from these diazotrophs would constitute a new paradigm for research on oceanic nitrogen cycling. Here we highlight the need for combining rate measurements and molecular analyses of field samples with cultivation studies in order to clarify the ecology of non-cyanobacteria and their contribution to marine N 2 fixation on local and global scales. [ABSTRACT FROM AUTHOR]

An initial modeling approach was applied to analyze how a single, nonmotile, free-living, heterotrophic bacterial cell may optimize the deployment of its extracellular enzymes. Free-living cells live in a dilute and complex substrate field, and to gain enough substrate, their extracellular enzymes must be utilized efficiently. The model revealed that surface-attached and free enzymes generate unique enzyme and substrate fields, and each deployment strategy has distinctive advantages. For a solitary cell, surface-attached enzymes are suggested to be the most cost-efficient strategy. This strategy entails potential substrates being reduced to very low concentrations. Free enzymes, on the other hand, generate a radically different substrate field, which suggests significant benefits for the strategy if free cells engage in social foraging or experience high substrate concentrations. Swimming has a slight positive effect for the attached-enzyme strategy, while the effect is negative for the free-enzyme strategy. The results of this study suggest that specific dissolved organic compounds in the ocean likely persist below a threshold concentration impervious to biological utilization. This could help explain the persistence and apparent refractory state of oceanic dissolved organic matter (DOM). Microbial extracellular enzyme strategies, therefore, have important implications for larger-scale processes, such as shaping the role of DOM in ocean carbon sequestration. [ABSTRACT FROM AUTHOR]

An initial modeling approach was applied to analyze how a single, nonmotile, free-living, heterotrophic bacterial cell may optimize the deployment of its extracellular enzymes. Free-living cells live in a dilute and complex substrate field, and to gain enough substrate, their extracellular enzymes must be utilized efficiently. The model revealed that surface-attached and free enzymes generate unique enzyme and substrate fields, and each deployment strategy has distinctive advantages. For a solitary cell, surface-attached enzymes are suggested to be the most cost-efficient strategy. This strategy entails potential substrates being reduced to very low concentrations. Free enzymes, on the other hand, generate a radically different substrate field, which suggests significant benefits for the strategy if free cells engage in social foraging or experience high substrate concentrations. Swimming has a slight positive effect for the attached-enzyme strategy, while the effect is negative for the free-enzyme strategy. The results of this study suggest that specific dissolved organic compounds in the ocean likely persist below a threshold concentration impervious to biological utilization. This could help explain the persistence and apparent refractory state of oceanic dissolved organic matter (DOM). Microbial extracellular enzyme strategies, therefore, have important implications for larger-scale processes, such as shaping the role of DOM in ocean carbon sequestration. [ABSTRACT FROM AUTHOR]

We investigated dinitrogen (N2) fixation activity and diazotroph community composition across the Cape Verde Frontal Zone (CVFZ), from photic epipelagic waters (0-200 m) to aphotic meso- (200–1000 m) and bathypelagic (> 1000 m) waters. The highest N2 fixation rates of 4.1 ± 2.2 nmol N l− 1 day− 1 and 7.8 ± 2.3 nmol N l− 1 day− 1 were in epipelagic waters south of the front. We detected aphotic N2 fixation in 5 out of 32 samples, primarily south of the front, and sporadically down to 3,000 m, with rates ranging from 0.03 ± 0.01 nmol N l− 1 day− 1 to 0.07 ± 0.01 nmol N l− 1 day− 1. Cyanobacteria dominated the diazotroph community and nitrogenase gene (nifH) expression profiles in surface waters and, surprisingly, in aphotic waters. The detection of cyanobacterial nifH genes by DNA sequencing and quantitative PCR in the aphotic zone, together with nifH expression in meso- and bathypelagic waters, indicates a downward flux of metabolically active cyanobacteria, and points to a contribution to the observed aphotic N2 fixation rates. In the photic zone, UCYN-A dominated north of the front, whereas Trichodesmium was mainly found in the southern region. However, our results also show that cross-frontal advection of cyanobacterial diazotrophs can occur via intrusions of surface water. Salinity, temperature, and mixed layer depth were the main determinants of the diazotroph composition and distribution of the key cyanobacteria. Thus, the front appeared to act as a dynamic barrier controlling the distribution of cyanobacterial diazotrophs. [ABSTRACT FROM AUTHOR]

In the Gulf of Bothnia, northern Baltic Sea, a large freshwater inflow creates north-southerly gradients in physico-chemical and biological factors across the two sub-basins, the Bothnian Bay (BB) and the Bothnian Sea. In particular, the sub-basins differ in nutrient limitation (nitrogen vs. phosphorus; P). Since viruses are rich in P, and virus production is commonly connected with bacterial abundance and growth, we hypothesized that the role of viral lysis differs between the sub-basins. Thus, we examined virus production and the potential importance of lysate recycling in surface waters along a transect in the Gulf of Bothnia. Surprisingly, virus production and total P were negatively correlated. In the BB, virus production rates were double those elsewhere in the system, although bacterial abundance and production were the lowest. In the BB, virus-mediated cell lysates could account for 70-180% and 100-250% of the bacterial carbon and P demand, respectively, while only 4-15% and 8-21% at the other stations. Low concentrations of dissolved DNA (D-DNA) with a high proportion of encapsulated DNA (viruses) in the BB suggested rapid turnover and high uptake of free DNA. The correlation of D-DNA and total P indicates that D-DNA is a particularly important nutrient source in the P-limited BB. Our study demonstrates large and counterintuitive differences in virus-mediated recycling of carbon and nutrients in two basins of the Gulf of Bothnia, which differ in microbial community composition and nutrient limitation. [ABSTRACT FROM AUTHOR]

Genetic, structural and physiological differences between strains of the marine bacterium Cellulophaga baltica MM#3 ( Flavobacteriaceae) developing in response to the activity of two virulent bacteriophages, ΦSM and ΦST, was investigated during 3 weeks incubation in chemostat cultures. A distinct strain succession towards increased phage resistance and a diversification of the metabolic properties was observed. During the incubation the bacterial population diversified from a single strain, which was sensitive to 24 tested Cellulophaga phages, into a multistrain and multiresistant population, where the dominant strains had lost susceptibility to up to 22 of the tested phages. By the end of the experiment the cultures reached a quasi steady state dominated by ΦST-resistant and ΦSM + ΦST-resistant strains coexisting with small populations of phage-sensitive strains sustaining both phages at densities of > 106 plaque forming units (pfu) ml−1. Loss of susceptibility to phage infection was associated with a reduction in the strains' ability to metabolize various carbon sources as demonstrated by BIOLOG assays. This suggested a cost of resistance in terms of reduced physiological capacity. However, there was no direct correlation between the degree of resistance and the loss of metabolic properties, suggesting either the occurrence of compensatory mutations in successful strains or that the cost of resistance in some strains was associated with properties not resolved by the BIOLOG assay. The study represents the first direct demonstration of phage-driven generation of functional diversity within a marine bacterial host population with significant implications for both phage susceptibility and physiological properties. We propose, therefore, that phage-mediated selection for resistant strains contributes significantly to the extensive microdiversity observed within specific bacterial species in marine environments. [ABSTRACT FROM AUTHOR]

Marine bacteria can cause harm to single-celled and multicellular eukaryotes. However, relatively little is known about the underlying genetic basis for marine bacterial interactions with higher organisms. We examined whole-genome sequences from a large number of marine bacteria for the prevalence of homologues to virulence genes and pathogenicity islands known from bacteria that are pathogenic to terrestrial animals and plants. As many as 60 out of 119 genomes of marine bacteria, with no known association to infectious disease, harboured genes of virulence-associated types III, IV, V and VI protein secretion systems. Type III secretion was relatively uncommon, while type IV was widespread among alphaproteobacteria (particularly among roseobacters) and type VI was primarily found among gammaproteobacteria. Other examples included homologues of the Yersinia murine toxin and a phage-related ‘antifeeding’ island. Analysis of the Global Ocean Sampling metagenomic data indicated that virulence genes were present in up to 8% of the planktonic bacteria, with highest values in productive waters. From a marine ecology perspective, expression of these widely distributed genes would indicate that some bacteria infect or even consume live cells, that is, generate a previously unrecognized flow of organic matter and nutrients directly from eukaryotes to bacteria. [ABSTRACT FROM AUTHOR]

Heterocystous filamentous cyanobacteria are regarded as the main N2-fixing organisms (diazotrophs) in the Baltic Sea. However, some studies indicate that picoplankton may also be important. The aim of this study was to examine the composition of putative diazotrophs in the picoplankton (< 3 µm) and to identify links to environmental factors. Nitrogenase ( nifH) genes were amplified from community DNA by nested PCR, followed by cloning and sequencing. Clone libraries from nine environmental samples collected from the central Baltic Sea (April–October 2003, 3 m depth) and a negative control yielded a total of 433 sequences with an average clone library coverage of 92%. The sequences fell within nifH Clusters I, II and III and formed 15 distinct groups (> 96% amino acid similarity). Most of the sequences (77%) fell into nifH Cluster I (cyanobacteria and α-, β- and γ- Proteobacteria). However, only 26 sequences were related to cyanobacteria (e.g. Pseudanabaena) and among these no unicellular phylotypes were found. Sequences clustering with alternative nitrogenases ( anfH) and Archaea were found in one sample while sequences related to anaerobic phylotypes were found in six samples distributed throughout the season. The identified phylogenetic groups showed covariance with several environmental factors but no strong links could be established. This suggests a variable and complex regulation of diazotrophic groups within Baltic Sea picoplankton. [ABSTRACT FROM AUTHOR]

Estuaries receive substantial anthropogenic nitrogen loading and are mainly considered net nitrogen sinks. While several studies have identified diverse diazotrophic communities in estuarine sediments, the role of pelagic diazotrophs in these systems is not well understood. We investigated the links between diazotrophic community composition, nitrogenase (nifH) gene expression, N2 fixation, and environmental conditions in Narragansett Bay (USA). Pelagic N2 fixation rates ranged between 0.02 and 9.41 nmol N L−1 d−1 and correlated significantly with fluctuations in diazotroph community composition. These fluctuations were also correlated with temperature, salinity, and mean sea level. The dominant sequences in our pelagic samples were related to sequences previously detected in the bay's sediments and were dominated by nifH gene Clusters I and III. We interpret this as a coupling between sediment and pelagic diazotroph communities and speculate that resuspension plays an important role for pelagic N2 fixation in shallow estuarine environments such as Narragansett Bay. For instance, the finding of active sulfate reducers in the oxygenated water illustrates that the sediment‐pelagic coupling can impact nutrient cycling in shallow environments. The pelagic N2 fixation measured during our study period showed only a minor contribution (< 1%) to the total estimated nitrogen load to Narragansett Bay. However, with intensifying nitrogen management in estuaries, the need to constrain the rates of pelagic N2 fixation in these systems will be essential for estimating nitrogen fluxes within the bay and to the adjacent coastal ecosystems. [ABSTRACT FROM AUTHOR]

The genome encodes the metabolic and functional capabilities of an organism and should be a major determinant of its ecological niche. Yet, it is unknown if the niche can be predicted directly from the genome. Here, we conduct metagenomic binning on 123 water samples spanning major environmental gradients of the Baltic Sea. The resulting 1961 metagenome-assembled genomes represent 352 species-level clusters that correspond to 1/3 of the metagenome sequences of the prokaryotic size-fraction. By using machine-learning, the placement of a genome cluster along various niche gradients (salinity level, depth, size-fraction) could be predicted based solely on its functional genes. The same approach predicted the genomes' placement in a virtual niche-space that captures the highest variation in distribution patterns. The predictions generally outperformed those inferred from phylogenetic information. Our study demonstrates a strong link between genome and ecological niche and provides a conceptual framework for predictive ecology based on genomic data. Alneberg et al. conduct metagenomics binning of water samples collected over major environmental gradients in the Baltic Sea. They use machine-learning to predict the placement of genome clusters along niche gradients based on the content of functional genes. [ABSTRACT FROM AUTHOR]

Temperature and CO2 levels are projected to increase in the future, with consequences for carbon and nutrient cycling in brackish environments, such as the Baltic Sea. Moreover, filamentous cyanobacteria are predicted to be favored over other phytoplankton groups under these conditions. Under a 12-day outdoor experiment, we examined the effect on a natural phytoplankton spring bloom community of elevated temperature (from 1°C to 4°C) and elevated pCO2 (from 390 to 970 μatm). No effects of elevated pCO2 or temperature were observed on phytoplankton biovolumes, but a significantly higher photosystem II activity was observed at elevated temperature after 9 days. In addition, three species of diazotrophic filamentous cyanobacteria were inoculated to test their competitive capacity under spring bloom conditions. The toxic cyanobacterium Nodularia spumigena exhibited an average specific growth rate of 0.10 d−1 by the end of the experiment, indicating potential prevalence even during wintertime in the Baltic Sea. Generally, none of the inoculated cyanobacteria species were able to outcompete the natural phytoplankton species at temperatures ≤4°C. No direct effects were found on heterotrophic bacteria. This study demonstrates the highly efficient resistance towards short-term (12 days) changes in abiotic factors by the natural Baltic Sea spring bloom community. [ABSTRACT FROM AUTHOR]

Highlights • This study describes the dinoflagellate and ciliate summer community in the Amundsen Sea Polynya. • Main taxa were mixotrophs and heterotrophs known to graze on Phaeocystis antarctica. • Other potential prey were also important in shaping the microzooplankton community. • Microscopy and sequencing results gave complementary insights on the community. Abstract In Antarctica, summer is a time of extreme environmental shifts resulting in large coastal phytoplankton blooms fueling the food web. Despite the importance of the microbial loop in remineralizing biomass from primary production, studies of how microzooplankton communities respond to such blooms in the Southern Ocean are rather scarce. Microzooplankton (ciliate and dinoflagellate) communities were investigated combining microscopy and 18S rRNA sequencing analyses in the Amundsen Sea Polynya during an extensive summer bloom of Phaeocystis antarctica. The succession of microzooplankton was further assessed during a 15-day induced bloom microcosm experiment. Dinoflagellates accounted for up to 59 % of the microzooplankton biomass in situ with Gymnodinium spp., Protoperidium spp. and Gyrodinium spp. constituting 89 % of the dinoflagellate biomass. Strobilidium spp., Strombidium spp. and tintinids represented 90 % of the ciliate biomass. Gymnodinium , Gyrodinium and tintinnids are known grazers of Phaeocystis, suggesting that this prymnesiophyte selected for the key microzooplankton taxa. Availability of other potential prey, such as diatoms, heterotrophic nanoflagellates and bacteria, also correlated to changes in microzooplankton community structure. Overall, both heterotrophy and mixotrophy appeared to be key trophic strategies of the dominant microzooplankton observed, suggesting that they influence carbon flow in the microbial food web through top-down control on the phytoplankton community. [ABSTRACT FROM AUTHOR]

Nitrogen (N 2) fixation rates in the brackish Baltic Sea are among the highest per unit of area in the world. However, beyond the filamentous heterocyst-forming cyanobacteria, knowledge about the composition and distribution of N 2 -fixing microbes (diazotrophs) is limited. To address this, we investigated nitrogenase gene (nifH) composition and expression at coastal (<10 km offshore) and offshore (>10 km offshore) stations, at surface (avg. 1.8 m) and at depth (avg. 24 m) and in free-living (0.2–3.0 μm) and particle-associated size fractions (>3 μm). Surprisingly, nifH genes affiliated with Pseudanabaena and non-cyanobacterial diazotrophs (NCDs) dominated the composition whereas filamentous heterocyst-forming cyanobacteria accounted for almost 80% of the nifH transcripts. Salinity had a minor influence on the composition, but Aphanizomenon and Nodularia showed increased relative nifH gene expression at low and higher salinity, respectively. Pseudanabaena only accounted for up to 5% of the nifH transcripts and nifH gene expression by Candidatus Atelocyanobacterium thalassa (sublineage UCYN-A2) was mainly observed in the most saline western part of the Baltic. The only notable expression by NCDs (up to 15% of nifH transcripts at a given station) coincided with an upwelling event at the southern coast and was largely accounted for by a Pseudomonas -like nifH phylotype, recurrently found in the Baltic Sea. NCD relative abundances were dominant in coastal stations, presumably driven by sediment resuspension as evidenced by higher turbidity and DOC levels and the recovery of sediment diazotrophs in the pelagic zone. This study reveals the heterogeneity of the composition and activity of diazotrophs in the Baltic Sea, and underscores the need for future N 2 fixation studies that include coastal and offshore Baltic waters. [Display omitted] • Pseudanabaena and non-cyanobacteria dominate the nitrogenase (nifH) gene pool. • Heterocyst-forming cyanobacteria accounted for 80% of the nifH gene transcripts. • Coastal upwelling affected composition and activity of diazotrophs. • The composition and activity of diazotrophs in the Baltic Sea is heterogeneous. • Future N 2 fixation studies should include coastal and offshore Baltic waters. [ABSTRACT FROM AUTHOR]

Vitamin B1 (B1 herein) is a vital enzyme cofactor required by virtually all cells, including bacterioplankton, which strongly influence aquatic biogeochemistry and productivity and modulate climate on Earth. Intriguingly, bacterioplankton can be de novo B1 synthesizers or B1 auxotrophs, which cannot synthesize B1 de novo and require exogenous B1 or B1 precursors to survive. Recent isolate-based work suggests select abundant bacterioplankton are B1 auxotrophs, but direct evidence of B1 auxotrophy among natural communities is scant. In addition, it is entirely unknown if bulk bacterioplankton growth is ever B1-limited. We show by surveying for B1-related genes in estuarine, marine, and freshwater metagenomes and metagenome-assembled genomes (MAGs) that most naturally occurring bacterioplankton are B1 auxotrophs. Pyrimidine B1-auxotrophic bacterioplankton numerically dominated metagenomes, but multiple other B1-auxotrophic types and distinct uptake and B1-salvaging strategies were also identified, including dual (pyrimidine and thiazole) and intact B1 auxotrophs that have received little prior consideration. Time-series metagenomes from the Baltic Sea revealed pronounced shifts in the prevalence of multiple B1-auxotrophic types and in the B1-uptake and B1-salvaging strategies over time. Complementarily, we documented B1/precursor limitation of bacterioplankton production in three of five nutrient-amendment experiments at the same time-series station, specifically when intact B1 concentrations were =3.7 pM, based on bioassays with a genetically engineered Vibrio anguillarum B1-auxotrophic strain. Collectively, the data presented highlight the prevalent reliance of bacterioplankton on exogenous B1/precursors and on the bioavailability of the micronutrients as an overlooked factor that could influence bacterioplankton growth and succession and thereby the cycling of nutrients and energy in aquatic systems. [ABSTRACT FROM AUTHOR]

Summary: Bacteria are major drivers of biogeochemical nutrient cycles and energy fluxes in marine environments, yet how bacterial communities respond to environmental change is not well known. Metagenomes allow examination of genetic responses of the entire microbial community to environmental change. However, it is challenging to link metagenomes directly to biogeochemical process rates. Here, we investigate metagenomic responses in natural bacterioplankton communities to simulated environmental stressors in the Baltic Sea, including increased river water input, increased nutrient concentration, and reduced oxygen level. This allowed us to identify informative prokaryotic gene markers, responding to environmental perturbation. Our results demonstrate that metagenomic and metabolic changes in bacterial communities in response to environmental stressors are influenced both by the initial community composition and by the biogeochemical factors shaping the functional response. Furthermore, the different sources of dissolved organic matter (DOM) had the largest impact on metagenomic blueprint. Most prominently, changes in DOM loads influenced specific transporter types reflecting the substrate availability and DOC assimilation and consumption pathways. The results provide new knowledge for developing models of ecosystem structure and biogeochemical cycling in future climate change scenarios and advance our exploration of the potential use of marine microorganisms as markers for environmental conditions. [ABSTRACT FROM AUTHOR]

Summary: Nitrification is important in nitrogen (N) cycling of aquatic environments, but knowledge about its regulation and importance is sparse. Here we examined nitrification and ammonia oxidizers in the Baltic Sea. We investigated two sites with different catchment characteristics (agricultural and forest), the Bay of Gdánsk (south) and the Öre Estuary (north), and measured pelagic nitrification rates and abundance, composition and expression of ammonia monooxygenase (amoA) genes. Highest nitrification rates were found in the nutrient rich Bay of Gdańsk. Interestingly, abundances of ammonia‐oxidizing archaea (AOA) and bacteria (AOB) were orders of magnitude lower than reported from other sites. Although AOA were most abundant at both sites, the highest expression levels were from AOB. Interestingly, few AOA and AOB taxa dominated amoA gene expression, with a Nitrosomarinus related phylotype showing widespread expression. AOA and AOB communities differed between sites and depths, respectively, with the composition in rivers being distinct. A storm event, causing an even depth distribution of nitrification and particles in the Bay of Gdańsk, indicated that the presence of particles stimulate nitrification. The study highlights coastal regions as dynamic sites of extensive pelagic nitrification, which may affect local food web dynamics and loss of N mediated by denitrification. [ABSTRACT FROM AUTHOR]

The European and American eels spawn in the subtropical convergence zone (STCZ) in the Sargasso Sea, a dynamic and relatively productive area that is strongly influenced by front and eddy formations and subducted high-saline water masses. To understand how the physical and biological environments may affect the early life history of eels, we conducted a detailed bio-physical investigation of the water column at a site of high eel larvae abundance. Diel measurements and sampling in the upper 300 m revealed strong variations in hydrographic conditions and mean depths of different taxonomic groups; however, characteristics patterns of distribution were apparent. Most species showed diel vertical migrations, ascending about 20–30 m at night, whereas examples of night–time downward migration were also seen. European eel larvae were among the species showing more extensive diel vertical migration: their population mean depth changed from 160 m at day to 100 m at night where abundance peaked at 45 m depth. Distribution and migration of eel larvae corresponded to patterns observed for small hydrozoans, supporting a proposed predator-prey linkage. The study demonstrates the diverse and vertically strongly structured plankton community of STCZ where larvae of eel and other fish find a wide range of potential niches. [ABSTRACT FROM AUTHOR]

Dissolved organic matter (DOM) in marine waters is a complex mixture of compounds and elements that contribute substantially to the global carbon cycle. The large reservoir of dissolved organic carbon (DOC) represents a vital resource for heterotrophic bacteria. Bacteria can utilise, produce, recycle and transform components of the DOM pool, and the physicochemical characteristics of this pool can directly influence bacterial activity; with consequences for nutrient cycling and primary productivity. In the present study we explored bacterial transformation of naturally occurring DOM across an extensive brackish water gradient in the Baltic Sea. Highest DOC utilisation (indicated by decreased DOC concentration) was recorded in the more saline southerly region where waters are characterised by more autochthonous DOM. These sites expressed the lowest bacterial growth efficiency (BGE), whereas in northerly regions, characterised by higher terrestrial and allochthonous DOM, the DOC utilisation was low and BGE was highest. Bacterial processing of the DOM pool in the south resulted in larger molecular weight compounds and compounds associated with secondary terrestrial humic matter being degraded, and a processed DOM pool that was more aromatic in nature and contributed more strongly to water colour; while the opposite was true in the north. Nutrient concentration and stoichiometry and DOM characteristics affected bacterial activity, including metabolic status (BGE), which influenced DOM transformations. Our study highlights dramatic differences in DOM characteristics and microbial carbon cycling in sub-basins of the Baltic Sea. These findings are critical for our understanding of carbon and nutrient biogeochemistry, particularly in light of climate change scenarios. [ABSTRACT FROM AUTHOR]

Production and bioavailability of dissolved organic matter (DOM) were followed during a year in the nutrient-rich estuary, Roskilde Fjord (RF), and the more oligotrophic strait, Great Belt (GB), in Denmark. Bioavailability of dissolved organic carbon (DOC), nitrogen (DON), and phosphorous (DOP) was determined during incubations over six months. Overall, RF had three to five times larger pools of total nitrogen (TN) and total phosphorous (TP) and five to eight times higher concentrations of inorganic nutrients compared to GB. However, the allocation of carbon, nitrogen, and phosphorous into different pools were remarkably similar between the two systems. DON and DOP contributed with about equal relative fractions in the two systems: 72 ± 13% of total nitrogen and 21 ± 12% of total phosphorous. The average bioavailability of DOM was 25 ± 15, 17 ± 5.5, and 49 ± 29% for carbon, nitrogen, and phosphorous, respectively. The observed release of DIN from degradation of DON amounted to between 0.1 (RF winter) and 14 times (GB summer) the loadings from land and contributed with half of the total input of bioavailable nitrogen during summer. Hence, this study shows that nitrogen in DOM is important for the nitrogen cycling, especially during summer. The sum of inorganic nutrients, particulate organic matter, and bioavailable DOM (the dynamic pools of nutrients) accounted for 42 and 92% of nitrogen, and phosphorous, respectively, and was remarkably similar between the two systems compared to the difference in nutrient richness. It is hypothesized that the pelagic metabolism of nutrients in marine systems dictates a rather uniform distribution of the different fractions of nitrogen and phosphorous containing compounds regardless of eutrophication level. [ABSTRACT FROM AUTHOR]

The number of icebergs produced from ice-shelf disintegration has increased over the past decade in Antarctica. These drifting icebergs mix the water column, influence stratification and nutrient condition, and can affect local productivity and food web composition. Data on whether icebergs affect bacterioplankton function and composition are scarce, however. We assessed the influence of iceberg drift on bacterial community composition and on their ability to exploit carbon substrates during summer in the coastal Southern Ocean. An elevated bacterial production and a different community composition were observed in iceberg-influenced waters relative to the undisturbed water column nearby. These major differences were confirmed in short-term incubations with bromodeoxyuridine followed by CARD-FISH. Furthermore, one-week bottle incubations amended with inorganic nutrients and carbon substrates (a mix of substrates, glutamine, N-acetylglucosamine, or pyruvate) revealed contrasting capacity of bacterioplankton to utilize specific carbon substrates in the iceberg-influenced waters compared with the undisturbed site. Our study demonstrates that the hydrographical perturbations introduced by a drifting iceberg can affect activity, composition, and substrate utilization capability of marine bacterioplankton. Consequently, in a context of global warming, increased frequency of drifting icebergs in polar regions holds the potential to affect carbon and nutrient biogeochemistry at local and possibly regional scales. [ABSTRACT FROM AUTHOR]

1. Planktonic bacterial community dynamics over short timescales can be of great importance for food webs and ecosystem functioning but are rarely described when microbial community and composition are assessed. To study the significance of such dynamics we sampled the surface water at the deepest point of a mesotrophic lake (Lake Erken, Sweden) every third hour over two days. 2. By combining 454 pyrosequencing of 16S rRNA genes with bromodeoxyuridine immunocapturing of DNA, replicating populations were identified and compared to the community retrieved from total DNA samples. This comparison revealed a significant difference between the actively replicating and total community. 3. The high-frequency diurnal sampling was compared to a year-long survey conducted in the same lake in order to compare the diurnal and seasonal variation in bacterioplankton community composition. At the diurnal-scale, the variation was significantly higher in the replicating than in the total community. However, variation in both active and total diurnal community was significantly lower than the variation in the seasonal total community. 4. Our analysis revealed pronounced short-term dynamics of individual bacterial populations uncoupled from the diurnal light cycle. For example, the proliferating fraction of the most abundant bacterial tribe (LD12) followed a cyclic pattern that covaried with viral abundance. This implies that environmental factors other than light may act as important drivers of microbial community composition, at least in mesotrophic Lake Erken. [ABSTRACT FROM AUTHOR]

Nitrogen (N 2) fixation by heterotrophic non-cyanobacterial diazotrophs is common in marine deep-sea sediments. However, in shallow coastal areas, where resuspension of sediments is extensive, the magnitude of sediment-associated N 2 fixation during resuspension is unknown. We examined nitrogen fixation in dark slurry incubations with sediments (0–5 cm and 0–10 cm depths) under simulated resuspension from five shallow stations (water depth <1 m) and an anoxic site in the Bay of Gdansk (109 m) in the Baltic Sea. Abiotic variables and the composition of nitrogen fixing organisms (diazotrophs) were measured at the study sites. To estimate the contribution of nitrogen fixing sulfate reducing bacteria to total nitrogen fixation, parallel incubations with sodium molybdate as inhibitor were performed. Our data show low but variable nitrogen fixation rates (n.d. - 23.7 nmol N g−1 d−1), promoted in small-grained sediments associated with increased organic carbon content and high nutrient concentrations in pore waters. Highest nitrogen fixation at the shallow sites was encountered in the upper 0–5 cm of the sediments while rates were negligible below. Sulfate reducing bacteria (e.g. Desulfobacterales and Desulfovibrionales) were responsible for most of the heterotrophic nitrogen fixation and appear as key players for pelagic N 2 fixation during resuspension. Our study reveals an important sediment – water coupling, which may be accentuated by the increased storms and resuspension events predicted for the Baltic Sea region. • Heterotrophic N 2 fixation occurs in shallow coastal sediments of the Baltic Sea. • N 2 fixation appears to be associated with small-grained sediments, possibly due to elevated levels of organic carbon and nutrients in the pore waters. • Diazotrophic SRB (Desulfobacterales and Desulfovibrio) perform most of dark N 2 fixation under simulated resuspension. [ABSTRACT FROM AUTHOR]

The community composition of N2-fixing microorganisms (diazotrophs) was investigated in copepods (primarily A cartia spp.) in parallel to that of seawater in coastal waters off Denmark ( Øresund) and New England, USA. The unicellular cyanobacterial diazotroph UCYN- A was detected from seawater and full-gut copepods, suggesting that the new N contributed by UCYN- A is directly transferred to higher trophic levels in these waters. Deltaproteobacterial and Cluster 3 nifH sequences were detected in > 1 μm seawater particles and full-gut copepods, suggesting that they associate with copepods primarily via feeding. The dominant communities in starved copepods were V ibrio spp. and related G ammaproteobacteria, suggesting they represent the most permanent diazotroph associations in the copepods. N2 fixation rates were up to 3.02 pmol N copepod−1 day−1. Although at a typical copepod density in estuarine waters, these volumetric rates are low; considering the small size of a copepod, these mesozooplanktonic crustaceans may serve as hotspots of N2 fixation, at 12.9-71.9 μmol N dm−3 copepod biomass day−1. Taken together, diazotroph associations range from more permanent attachments to copepod feeding on some groups. Similar diazotroph groups detected on the eastern and western Atlantic Ocean suggest that these associations are a general phenomenon and play a role in the coastal N cycles. [ABSTRACT FROM AUTHOR]

Due to climate change, sea ice experiences changes in terms of extent and physical properties. In order to understand how sea ice microbial communities are affected by changes in physicochemical properties of the ice, we used 454-sequencing of 16 S and 18 S rRNA genes to examine environmental control of microbial diversity and composition in Antarctic sea ice. We observed a high diversity and richness of bacteria, which were strongly negatively correlated with temperature and positively with brine salinity. We suggest that bacterial diversity in sea ice is mainly controlled by physicochemical properties of the ice, such as temperature and salinity, and that sea ice bacterial communities are sensitive to seasonal and environmental changes. For the first time in Antarctic interior sea ice, we observed a strong eukaryotic dominance of the dinoflagellate phylotype SL163 A10, comprising 63% of the total sequences. This phylotype is known to be kleptoplastic and could be a significant primary producer in sea ice. We conclude that mixotrophic flagellates may play a greater role in the sea ice microbial ecosystem than previously believed, and not only during the polar night but also during summer when potential food sources are abundant. [ABSTRACT FROM AUTHOR]

Jellyfish are effective predators on mesozooplankton and release large amounts of dissolved organic matter. Nevertheless, jellyfish initiated trophic cascades and bottom-up influences impacting lower trophic levels have received limited attention. We conducted a mesocosm experiment to quantify simultaneous top-down and bottom-up effects of a common jellyfish, Cyanea capillata, in a natural plankton community during autumn. Treatments were 0, 2 or 5 jellyfish per 2.5 m³ mesocosm, four replicates each, with initial additions of inorganic nutrients. Primary and bacterial production, species abundance and composition of several trophic levels and nutrient and carbon dynamics were followed during the 8-day experiment. Multivariate statistics and generalized additive mixed modelling were applied to test whether jellyfish carbon concentration (0-1.26 mg jellyC L-1) in the mesocosms affected the variables monitored. Unexpected negligible predatory impact of jellyfish on mesozooplankton was observed, potentially related to jellyfish senescence. Community compositions of bacteria, phytoplankton and mesozooplankton changed with time, but did not differ between treatments. However, nutrient regeneration by jellyfish was evident, and jellyfish had a positive impact on total and specific bacterial production, total primary production and the >10 μm chlorophyll a fraction. Bottom-up influences from abundant jellyfish could thus stimulate productivity in nutrient depleted autumnal surface waters. [ABSTRACT FROM AUTHOR]

Microbial communities associated with marine sponges carry out nutrient transformations essential for benthic-pelagic coupling; however, knowledge about their composition and function is still sparse. We evaluated the richness and diversity of prokaryotic assemblages associated with three high-microbial-abundance (HMA) and three low-microbial-abundance (LMA) sympatric Mediterranean sponges to address their stability and uniqueness. Moreover, to examine functionality and because an imbalance between nitrogen ingestion and excretion has been observed for some of these species, we sequenced nitrogenase genes (nifH) and measured N2 fixation. The prokaryotic communities in the two sponge types did not differ in terms of richness, but the highest diversity was found in HMA sponges. Moreover, the discrete composition of the communities in the two sponge types relative to that in the surrounding seawater indicated that horizontal transmission and vertical transmission affect the microbiomes associated with the two sponge categories. nifH genes were found in all LMA species and sporadically in one HMA species, and about half of the nifH gene sequences were common between the different sponge species and were also found in the surrounding water, suggesting horizontal transmission. 15N2-enriched incubations showed that N2 fixation was measurable in the water but was not associated with the sponges. Also, the analysis of the isotopic ratio of 15N to 14N in sponge tissue indicated that N2 fixation is not an important source of nitrogen in these Mediterranean sponges. Overall, our results suggest that compositional and functional features differ between the prokaryotic communities associated with HMA and LMA sponges, which may affect sponge ecology. [ABSTRACT FROM AUTHOR]

The genetic capacity to fix gaseous nitrogen (N) is distributed among diverse diazotrophs belonging to the Bacteria and Archaea. However, only a subset of the putative diazotrophs present actively fix N at any given time in the environment. We experimentally tested whether the availability of carbon and inhibition by oxygen constrain N fixation by diazotrophs in coastal seawater. The goal was to test whether by alleviating these constraints an increased overlap between nitrogenase (nifH)-gene-carrying and -expressing organisms could be achieved. We incubated water from a eutrophic but N-limited fjord in Denmark under high-carbon/low-oxygen conditions and determined bacterial growth and production, diazotrophic community composition (Illumina nifH amplicon sequencing), and nifH gene abundance and expression [quantitative PCR (qPCR) and quantitative reverse transcriptase PCR (qRT-PCR)]. Bacterial abundances and production increased under high-carbon/low-oxygen conditions as did the similarity between present and active diazotrophic communities. This was caused by the loss of specific abundant yet non-active gammaproteobacterial phylotypes and increased expression by others. The prominent active gamma- and epsilonproteobacterial diazotrophs did not, however, respond to these conditions in a uniform way, highlighting the difficulty to assess how a change in environmental conditions may affect a diverse indigenous diazotrophic community. [ABSTRACT FROM AUTHOR]

Nitrogen (N) fixation is fueling planktonic production in a multitude of aquatic environments. In meso- and poly-haline estuaries, however, the contribution of N by pelagic N2 fixation is believed to be insignificant due to the high input of N from land and the presumed absence of active N2-fixing organisms. Here we report N2 fixation rates, nifH gene composition and nifH gene transcript abundance for key diazotrophic groups over 1 year in two contrasting, temperate, estuarine systems: Roskilde Fjord (RF) and the Great Belt (GB) strait. Annual pelagic N2 fixation rates averaged 17 and 61 mmol N m−2 per year at the two sites, respectively. In RF, N2 fixation was mainly accompanied by transcripts related to heterotrophic (for example, Pseudomonas sp.) and photoheterotrophic bacteria (for example, unicellular diazotrophic cyanobacteria group A). In the GB, the first of two N2 fixation peaks coincided with a similar nifH-expressing community as in RF, whereas the second peak was synchronous with increased nifH expression by an array of diazotrophs, including heterotrophic organisms as well as the heterocystous cyanobacterium Anabaena. Thus, we show for the first time that significant planktonic N2 fixation takes place in mesohaline, temperate estuaries and that the importance of heterotrophic, photoheterotrophic and photosynthetic diazotrophs is clearly variable in space and time. [ABSTRACT FROM AUTHOR]

The wide distribution of diverse nitrogenase ( nifH) genes affiliated with those of heterotrophic bacteria in marine and estuarine waters indicates ubiquity and an ecologically relevant role for heterotrophic N2-fixers (diazotrophs) in aquatic nitrogen ( N) cycling. However, the lack of cultivated representatives currently precludes an evaluation of their N2-fixing capacity. In this study, microoxic or anoxic N-free media were inoculated with estuarine Baltic Sea surface water to select for N2-fixers. After visible growth and isolation of single colonies on oxic plates or in anoxic agar tubes, nifH gene amplicons were obtained from 64 strains and nitrogenase activity, applying the acetylene reduction assay, was confirmed for 40 strains. Two strains, one G ammaproteobacterium affiliated with P seudomonas and one A lphaproteobacterium affiliated with R hodopseudomonas were shown to represent established members of the indigenous diazotrophic community in the Baltic Sea, with abundances of up to 7.9 × 104 and 4.7 × 104 nifH copies l−1 respectively. This study reports media for successful isolation of heterotrophic diazotrophs. The applied methodology and the obtained strains will facilitate future identification of factors controlling heterotrophic diazotrophic activity in aquatic environments, which is a prerequisite for understanding and evaluating their ecology and contribution to N cycling at local and regional scales. [ABSTRACT FROM AUTHOR]

To explore the potential linkage between distribution of marine bacterioplankton groups, environmental conditions, and water mass, we investigated the factors determining the abundance of bacterial taxa across the hydrographically complex Subtropical Convergence Zone in the Sargasso Sea. Based on information from 16S rRNA gene clone libraries from various locations and two depths, abundances of the predominant taxa (eubacteria, Archaea, Alphaproteobacteria, Gatnmaproteobacteria, Bacteroidetes, and the Roseobacter, SARI 1, and SAR86 clades) were quantified by real-time PCR. In addition, the abundances of Synechococcus, Prochlorococcus, and picoalgae were determined by flow cytometry. Linear multiple-regression models determining the relative effects of eight environmental variables and of water mass explained 35 to 86% of the variation in abundance of the quantified taxa, even though only one to three variables were significantly related to any particular taxon's abundance. Most of the variation in abundance was explained by depth and chlorophyll a. The predominant phototrophs, Prochlorococcus and picoalgae, were negatively correlated with phosphate, whereas eubacteria, heterotrophic bacteria, and SAR86 were negatively correlated with nitrite. Water mass showed limited importance for explaining the abundance of the taxonomical groups (significant only for Roseobacter, explaining 14% of the variation). The results suggest the potential for predicting the abundance of broad bacterioplankton groups throughout the Sargasso Sea using only a few environmental parameters. [ABSTRACT FROM AUTHOR]

Nitrogenase genes ( nifH) from heterotrophic dinitrogen (N2)-fixing bacteria appear ubiquitous in marine bacterioplankton, but the significance of these bacteria for N cycling is unknown. Quantitative data on the N2-fixation potential of marine and estuarine heterotrophs are scarce, and the shortage of cultivated specimens currently precludes ecophysiological characterization of these bacteria. Through the cultivation of diazotrophs from suboxic (1.79 μmol O2 L−1) Baltic Sea water in an artificial seawater medium devoid of combined N, we report the cultivability of a considerable fraction of the diazotrophic community in the Gotland Deep. Two nifH clades were present both in situ and in enrichment cultures showing gene abundances of up to 4.6 × 105 and 5.8 × 105 nifH gene copies L−1 within two vertical profiles in the Baltic Sea. The distributions of the two clades suggested a relationship with the O2 concentrations in the water column as abundances increased in the suboxic and anoxic waters. It was possible to cultivate and isolate representatives from one of these prevalent clades, and preliminary analysis of their ecophysiology demonstrated growth optima at 0.5-15 μmol O2 L−1 and 186-194 μmol O2 L−1 in the absence of combined N. [ABSTRACT FROM AUTHOR]

Studies indicate that bacterial taxa utilize different fractions of the dissolved organic carbon ( DOC) pool, while others suggest functional redundancy among constituents of bacterioplankton, implying only a weak coupling between community structure and function. We examined bacterial compositional and functional [ectoenzymatic activities and growth efficiency; bacterial growth efficiency ( BGE)] responses to a gradient in bioavailable DOC ( bDOC). This was achieved over 10 days in DOC utilization assays containing Baltic Sea water with variable amounts of natural bDOC. Measurements of bacterial growth, O2 and DOC consumption in the assays using non-invasive sampling showed that BGE changed over time and that the bDOC utilized accounted for 4-13% of the DOC pool. Pyrosequencing of 16S rRNA genes demonstrated minor differences at the phylum level between samples, whereas larger successional differences were discernible at lower phylogenetic levels. Our study suggests that changes in concentrations of bDOC affect bacterioplankton BGE and community structure by selecting for some taxa while the relative abundance of most taxa remained unaffected. Ectoenzymes activities suggested preferential degradation of protein-rich compounds by bacteria, switching to carbohydrate-rich DOC when proteins were depleted. Hence, there was a fairly weak linkage between bacterial community composition and DOC utilization suggesting that overall bacterioplankton community structure only to some extent has predictive power for processing of the DOC pool. [ABSTRACT FROM AUTHOR]

Viruses are fundamental to ecosystems ranging from oceans to humans, yet our ability to study them is bottlenecked by the lack of ecologically relevant isolates, resulting in "unknowns" dominating culture-independent surveys. Here we present genomes from 31 phages infecting multiple strains of the aquatic bacterium Cellulophaga baltica (Bacteroidetes) to provide data for an under-represented and environmentally abundant bacterial lineage. Comparative genomics delineated 12 phage groups that (i) each represent a new genus, and (ii) represent one novel and four well-known viral families. This diversity contrasts the few well-studied marine phage systems, but parallels the diversity of phages infecting human-associated bacteria. Although all 12 Cellulophaga phages represent new genera, the podoviruses and icosahedral, nontailed ssDNA phages were exceptional, with genomes up to twice as large as those previously observed for each phage type. Structural novelty was also substantial, requiring experimental phage proteomics to identify 83% of the structural proteins. The presence of uncommon nucleotide metabolism genes in four genera likely underscores the importance of scavenging nutrient-rich molecules as previously seen for phages in marine environments. Metagenomic recruitment analyses suggest that these particular Cellulophaga phages are rare and may represent a first glimpse into the phage side of the rare biosphere. However, these analyses also revealed that these phage genera are widespread, occurring in 94% of 137 investigated metagenomes. Together, this diverse and novel collection of phages identifies a small but ubiquitous fraction of unknown marine viral diversity and provides numerous environmentally relevant phage-host systems for experimental hypothesis testing. [ABSTRACT FROM AUTHOR]