Factors controlling the community composition of marine heterotrophic prokaryotes include organic-C, mineral nutrients, predation, and viral lysis. Two mesocosm experiments, performed at an Arctic location and bottom-up manipulated with organic-C, had very different results in community composition for both prokaryotes and viruses. Previously, we showed how a simple mathematical model could reproduce food web level dynamics observed in these mesocosms, demonstrating strong top-down control through the predator chain from copepods via ciliates and heterotrophic nanoflagellates. Here, we use a steady-state analysis to connect ciliate biomass to bacterial carbon demand. This gives a coupling of top-down and bottom-up factors whereby low initial densities of ciliates are associated with mineral nutrient-limited heterotrophic prokaryotes that do not respond to external supply of labile organic-C. In contrast, high initial densities of ciliates give carbon-limited growth and high responsiveness to organic-C. The differences observed in ciliate abundance, and in prokaryote abundance and community composition in the two experiments were in accordance with these predictions. Responsiveness in the viral community followed a pattern similar to that of prokaryotes. Our study provides a unique link between the structure of the predator chain in the microbial food web and viral abundance and diversity.

The study of airborne bacteria relies on a sampling strategy that preserves their integrity and in situ physiological state, e.g. viability, cultivability, metabolic activity, and ice-nucleation activity. Because ambient air harbors low concentrations of bacteria, an effective bioaerosol sampler should have a high sampling efficiency and a high airflow. We characterize a high-flow-rate impinger with respect to particle collection and retention efficiencies in the range 0.5-3.0 μm, and we investigated its ability to preserve the physiological state of selected bacterial species and seawater bacterial community in comparison with four commercial bioaerosol samplers. The collection efficiency increased with particle size and the cutoff diameter was between 0.5 and 1 μm. During sampling periods of 120-300 min, the impinger retained the cultivability, metabolic activity, viability, and ice-nucleation activity of investigated bacteria. Field studies in semiurban, high-altitude, and polar environments included periods of low bacterial air concentrations, thus demonstrating the benefits of the impinger's high flow rate. In conclusion, the impinger described here has many advantages compared with other bioaerosol samplers currently on the market: a potential for long sampling time, a high flow rate, a high sampling and retention efficiency, low costs, and applicability for diverse downstream microbiological and molecular analyses.

P-transport through the microbial food web was investigated in a mesocosm experiment in which orthophosphate was added to oligotrophic Cretan Sea water. As non-exclusive alternatives to traditional phytoplankton-zooplankton succession, two mechanisms for P-transport have previously been proposed: a stoichiometric shift in food quality (Tunnelling); and a transport through a predatory food chain from heterotrophic bacteria, bypassing phytoplankton (Bypass). Following P-addition, particulate C:P-ratio dropped from 436 to 44(molar) within 1 day, and egg production increased after 2 days. This confirms the hypothesized stoichiometric shift and rapid copepod response of a "Tunnelling" scenario. Bacterial abundance responded positively to P-addition on Day 1, ciliates increased after Day 5 and new egg production peaks occurred on Days 5 and 9; a succession suggesting additional Bypass transport although the response expected in heterotrophic flagellate abundance was not confirmed. A small, but statistically not significant, increase in Chl a in the 0.6- to 2-mm size fraction also suggests possible additional P-transport through a phytoplankton zooplankton succession. On the basis of the magnitude of the stoichiometric change and the short delay in egg production response, we consider the Tunnelling mechanism to be the most likely signal initiating egg production, but a Bypass, and possibly a traditional succession, may have contributed to maintain the elevated egg production.

A minimum mathematical model of the marine pelagic microbial food web has previously shown to be able to reproduce central aspects of observed system response to different bottom-up manipulations in a mesocosm experiment Microbial Ecosystem Dynamics (MEDEA) in Danish waters. In this study, we apply this model to two mesocosm experiments (Polar Aquatic Microbial Ecology (PAME)-I and PAME-II) conducted at the Arctic location Kongsfjorden, Svalbard. The different responses of the microbial community to similar nutrient manipulation in the three mesocosm experiments may be described as diatom-dominated (MEDEA), bacteria-dominated (PAME-I), and flagellated-dominated (PAME-II). When allowing ciliates to be able to feed on small diatoms, the model describing the diatom-dominated MEDEA experiment give a bacteria-dominated response as observed in PAME I in which the diatom community comprised almost exclusively small-sized cells. Introducing a high initial mesozooplankton stock as observed in PAME-II, the model gives a flagellate-dominated response in accordance with the observed response also of this experiment. The ability of the model originally developed for temperate waters to reproduce population dynamics in a 10°C colder Arctic fjord, does not support the existence of important shifts in population balances over this temperature range. Rather, it suggests a quite resilient microbial food web when adapted to in situ temperature. The sensitivity of the model response to its mesozooplankton component suggests, however, that the seasonal vertical migration of Arctic copepods may be a strong forcing factor on Arctic microbial food webs.

The mechanisms by which phytoplankton cope with stressors in the marine environment are neither fully characterized nor understood. As viruses are the most abundant entities in the global ocean and represent a strong top-down regulator of phytoplankton abundance and diversity, we sought to characterize the cellular response of two marine haptophytes to virus infection in order to gain more knowledge about the nature and diversity of microalgal responses to this chronic biotic stressor. We infected laboratory cultures of the haptophytes Haptolina ericina and Phaeocystis pouchetii with CeV-01B or PpV-01B dsDNA viruses, respectively, and assessed the extent to which host cellular responses resemble programmed cell death (PCD) through the activation of diagnostic molecular and biochemical markers. Pronounced DNA fragmentation and activation of cysteine aspartate-specific proteases (caspases) were only detected in virus-infected cultures of these phytoplankton. Inhibition of host caspase activity by addition of the pan-caspase inhibitor z-VAD-fmk did not impair virus production in either host-virus system, differentiating it from the Emiliania huxleyi-Coccolithovirus model of haptophyte-virus interactions. Nonetheless, our findings point to a general conservation of PCD-like activation during virus infection in ecologically diverse haptophytes, with the subtle heterogeneity of cell death biochemical responses possibly exerting differential regulation on phytoplankton abundance and diversity.

Storm clouds frequently form in the summer period in temperate climate zones. Studies on these inaccessible and short-lived atmospheric habitats have been scarce. We report here on the first comprehensive biogeochemical investigation of a storm cloud using hailstones as a natural stochastic sampling tool. A detailed molecular analysis of the dissolved organic matter in individual hailstones via ultra-high resolution mass spectrometry revealed the molecular formulae of almost 3000 different compounds. Only a small fraction of these compounds were rapidly biodegradable carbohydrates and lipids, suitable for microbial consumption during the lifetime of cloud droplets. However, as the cloud environment was characterized by a low bacterial density (Me = 1973 cells/ml) as well as high concentrations of both dissolved organic carbon (Me = 179 µM) and total dissolved nitrogen (Me = 30 µM), already trace amounts of easily degradable organic compounds suffice to support bacterial growth. The molecular fingerprints revealed a mainly soil origin of dissolved organic matter and a minor contribution of plant-surface compounds. In contrast, both the total and the cultivable bacterial community were skewed by bacterial groups (γ-Proteobacteria, Sphingobacteriales and Methylobacterium) that indicated the dominance of plant-surface bacteria. The enrichment of plant-associated bacterial groups points at a selection process of microbial genera in the course of cloud formation, which could affect the long-distance transport and spatial distribution of bacteria on Earth. Based on our results we hypothesize that plant-associated bacteria were more likely than soil bacteria (i) to survive the airborne state due to adaptations to life in the phyllosphere, which in many respects matches the demands encountered in the atmosphere and (ii) to grow on the suitable fraction of dissolved organic matter in clouds due to their ecological strategy. We conclude that storm clouds are among the most extreme habitats on Earth, where microbial life exists.

The atmosphere is host to an omnipresent bacterial community that may influence fundamental atmospheric processes such as cloud formation and precipitation onset. Knowledge of this bacterial community is scarce, particularly in air masses relevant to cloud formation. Using a light aircraft, we sampled above the atmospheric boundary layer—that is, at heights at which cloud condensation occurs—over coastal areas of Sweden and Denmark in summer 2009. Enumeration indicated total bacterial numbers of 4 × 101 to 1.8 × 103 m−3 air and colony-forming units of 0–6 bacteria m−3 air. 16S rRNA gene libraries constructed from samples collected above the Baltic Sea coast revealed a highly diverse bacterial community dominated by species belonging to the genera Sphingomonas and Pseudomonas. Bacterial species known to carry ice-nucleating proteins were found in several samples. Modeled back trajectories suggested the potential sources of the sampled bacteria to be diverse geographic regions, including both marine and terrestrial environments in the northern hemisphere. Several samples contained 16S rRNA genes from plant chloroplasts, confirming a terrestrial contribution to these samples. Interestingly, the airborne bacterial community displayed an apparent seasonal succession that we tentatively ascribe to in situ succession in the atmosphere.

Ocean acidification may stimulate primary production through increased availability of inorganic carbon in the photic zone, which may in turn change the biogenic flux of dissolved organic carbon (DOC) and the growth potential of heterotrophic bacteria. To investigate the effects of ocean acidification on marine bacterial assemblages, a two-by-three factorial mescosom experiment was conducted using surface sea water from the East Greenland Current in Fram Strait. Pyrosequencing of the V1-V2 region of bacterial 16S ribosomal RNA genes was used to investigate differences in the endpoint (Day 9) composition of bacterial assemblages in mineral nutrient-replete mesocosms amended with glucose (0 μM, 5.3 μM and 15.9 μM) under ambient (250 μatm) or acidified (400 μatm) partial pressures of CO(2) (pCO(2)). All mesocosms showed low richness and diversity by Chao1 estimator and Shannon index, respectively, with general dominance by Gammaproteobacteria and Flavobacteria. Nonmetric multidimensional scaling analysis and two-way analysis of variance of the Jaccard dissimilarity matrix (97% similarity cut-off) demonstrated that the significant community shift between 0 μM and 15.9 μM glucose addition at 250 μatm pCO(2) was eliminated at 400 μatm pCO(2). These results suggest that the response potential of marine bacteria to DOC input may be altered under acidified conditions.

The purpose of the present study was to explore the composition and variation of the pico-, nano- and micro-plankton communities in Norwegian coastal waters and Skagerrak, and the co-occurrence of bacteria and viruses. Samples were collected along three cruise transects from Jæren, Lista and Oksøy on the south coast of Norway and into the North Sea and Skagerrak. We also followed a drifting buoy for 55h in Skagerrak in order to observe diel variations. Satellite ocean color images (SeaWiFS) of the chlorophyll a (chl a) distribution compared favorably to in situ measurements in open waters, while closer to the shore remote sensing chl a data was overestimated compared to the in situ data. Using light microscopy, we identified 49 micro- and 15 nanoplankton sized phototrophic forms as well as 40 micro- and 12 nanoplankton sized heterotrophic forms. The only picoeukaryote (0.2–2.0μm) we identified was Resultor micron (Pedinophyceae). Along the transects a significant variation in the distribution and abundance of different plankton forms were observed, with Synechococcus spp and autotrophic picoeukaryotes as the most notable examples. There was no correlation between viruses and chl a, but between viruses and bacteria, and between viruses and some of the phytoplankton groups, especially the picoeukaryotes. Moreover, there was a negative correlation between nutrients and small viruses (Low Fluorescent Viruses) but a positive correlation between nutrients and large viruses (High Fluorescent Viruses). The abundance of autotrophic picoplankton, bacteria and viruses showed a diel variation in surface waters with higher values around noon and late at night and lower values in the evening. Synechococcus spp were found at 20m depth 25–45 nautical miles from shore apparently forming a bloom that stretched out for more than 100 nautical miles from Skagerrak and up the south west coast of Norway. The different methods used for assessing abundance, distribution and diversity of microorganisms yielded complementary information about the plankton community. Flow cytometry enabled us to map the distribution of the smaller phytoplankton forms, bacteria and viruses in more detail than has been possible before but detection and quantification of specific forms (genus or species) still requires taxonomic skills, molecular analysis or both.

Marine microbial communities have been little studied in Arctic waters, especially during the winter spring transition before the development of extensive phytoplankton blooms. This study investigated microbial plankton in the ice-covered polar surface waters of the northwestern Fram Strait (75 to 80 degrees N) at the onset of the 24 h light period in spring (April to May). The system we encountered was characterised by low concentrations of chlorophyll a (< 0.2 mu g l(-1)) and a low abundance of both bacteria (1.4 to 2.5 x 10(8) cells l(-1)) and protists (1 to 1.7 x 10(5) cells l(-1)). Bacterial production was very low (<= 0.63 mu g C l(-1) d(-1)), despite the dominance of nucleic-acid-rich bacteria (58 +/- 6% of total bacterial abundance). Small (2 to 5 mu m) phototrophs dominated the eukaryotic assemblage in the surface and most probably had profound effects on the composition and metabolic balance of the microbial community as a whole. Most stations appeared to have been net-autotrophic, and calculations of phagotrophy indicated a balanced carbon budget for the microbial community. Mixotrophy was seen in a large part of the ciliate assemblage and may have contributed to the productivity and stability of the pre-bloom system that we encountered.

The effect of water treatment and flow rate on young Atlantic cod juveniles was investigated in a 36-days experiment. Four different flow rates (10, 20, 40, and 70 times the effective tank volume per day) were set up in triplicate tanks within each of three rigs with recirculated, UV-radiated, and untreated water, respectively. Each of the 36 tanks was stocked with 200 weaned cod juveniles at a mean weight of 0.048g. Fish mortality was recorded daily in all tanks, and growth (wet weight) was determined at the end of the experiment. The microflora in the rearing water was investigated by means of PCR-DGGE and flow cytometry. Observed mortality was significantly higher at low flow rates while otherwise unexplained mortality (presumed to be due to cannibalism) was lowest in the recirculation system. No correlation was found between survival and growth. Growth was significantly affected by both water exchange rate and treatment, as the juveniles from high flow rates and the UV-treatment showed elevated growth rates. Both growth and survival scaled in accordance with metabolic factors like oxygen saturation and unionized ammonia. Bacterial concentrations increased in all tanks and treatment from the beginning of the experiment to the end. The UV-treated and untreated water started at typical seawater concentrations (0.5–1×106mL−1) and increased five to tenfold during the experiment. The recycled water tanks started with bacterial concentrations 2–5 times higher than the UV- and untreated experiments at the time of fish transfer, and ended up with 10 times higher concentrations in the end. Cluster analysis of the DGGE profiles separated the recirculation tanks, including the respective inlet water, from the flow-through systems, with one exception (the highest flow rate). Eighty-five% of the sequences clustered within the Gammaproteobacteria, further divided into four distinct clusters. One of the clusters was only detected in the recirculation system, and showed highest affiliation to bacteria belonging to the Alteromonas/Pseudoalteromonas genera. In contrast, bacteria belonging to the family Vibrionaceae were detected in the flow-through systems.

Bacteria are ubiquitous in the atmosphere, where they form a highly diverse community, albeit low in abundance. Several approaches are available for collecting airborne particles, though few comparative studies have been conducted to date. This study examined how different sampling strategies affect the apparent composition of the airborne community. Three devices were tested: an impactor, a liquid impinger, and a Teflon membrane filter. Comparative studies were conducted at one mountainous location in Norway and one seaside location in Sweden. At both locations, microbial samples were collected in parallel using the sampling devices. DNA extraction, construction of 16S rRNA gene clone libraries, and subsequent sequencing were used to identify the bacteria. The comparison between clone libraries retrieved using the different devices indicated good agreement regarding dominant species, overall diversity, and distribution of species among phylogenetic groups. Among the less common species, there were few shared sequences in different clone libraries, likely due to the high diversity of the assessed samples. Bacteria belonging to the Bacteroidetes and Proteobacteria phyla dominated at both locations, and the most common genera were Sphingomonas sp. and Pantoea sp. Chloroplast-like 16S rRNA gene sequences were detected in all samples.

To investigate the influences of viral infection on cell cycle of marine algae, we examined the changes of expression and activity of G(2)/M-phase cell cycle regulators in two unicellular marine algae Emiliania huxleyi and Chrysochromulina ericina after viral infection by immunoblot. The results revealed that the expression of cyclins could be altered by viral infection, thus inhibiting the progression of cell cycle. E. huxleyi virus (EhV) repressed the activity of p34cdc2/cyclinB complex by inactivating p34cdc2 kinase through phosphorylation of Cdc2, and block the life cycle of host cells at G(2)/M checkpoint. C. ericina virus (CeV) interfere with the activity of p34cdc2/cyclin B complex by the downregulation of cyclinB combined with the overexpression of kinase-active p34cdc2, resulting in a G(2) cell cycle. The G(2)/M cell cycle arrest caused by EhV infection indicated that EhV may transcribe its own genes without relying on the host cell cycle control. C. ericina cells blocked in a delayed G(2) phase may partly turn into the second S phase to synthesize proteins useful for viral nucleic acid metabolism, suggesting that the replication of CeV viruses was dependent on cell cycle control. Our results indicated that algal virus infection selectively activates/inactivates certain components of the cell cycle with the aim to establish a more efficient environment for their gene expression and DNA replication.

The authors have investigated the biochemical events by which marine algal virus infection induces cell cycle arrest. The key G(2)/M-phase regulatory proteins are analyzed by immunobloting in unicellular Emiliania huxleyi, suggesting that virus induced cell cycle arrest is related with virus's effect on cyclins and cyclin dependent kinases. E. huxleyi virus (EhV) represses Cdc2/cyclinB complex activity by inhibiting the activity of Cdc2 kinase in a phosphorylation-related manner, blocking host cells G(2)/M checkpoint. Dephosphorylated / inactive Cdc25C combined with up-regulation of Wee1 expression at early infect period appears to be important mechanisms by which EhV represses Cdc2/cyclinB complex activity that is required for entry into M phase. This study has allowed us to confirm that algal virus infection leads to selective activation or inhibition of certain cell-cycle factors, which may play a significant role in establishing a more efficient environment for viral gene expression and DNA replication.

In order to study interactions between microorganisms at different nutrient conditions in an arctic environment, a mesocosm experiment was performed in Kongsfjorden, Svalbard (79°N). A phytoplankton bloom was initiated by daily additions of mineral nutrients (ammonium and phosphate) to all mesocosm units. The addition of silicate and glucose, forming a factorial design (+Si/+C, +Si/−C, −Si/+C, −Si/−C), was intended to produce different types of growth rate limitation for the bacterial community. We here focus on the response in bacterial community composition to different nutrient situations. Phytoplankton, bacteria and viruses were enumerated by flow cytometry, while denaturing gradient gel electrophoresis (DGGE) was used to track changes in the bacterial community composition. Our results showed that both glucose and silicate addition affected the bacterial community composition, with the largest effect from glucose. The initial increase in bacterial abundance was most pronounced in the glucose units. After silicate addition, highest bacterial abundance was observed in the silicate treatments where mineral nutrient competition by diatoms was expected to be highest. The major effect of glucose was expressed by the significant separation of the +C and the −C samples at the end of the experiment, while silicate addition resulted in a more stable bacterial community structure. In the unit, given both silicate and glucose, the diatoms were totally outcompeted by the bacterial community. The competitive success of the heterotrophic bacteria in C-replete situations allows the conclusion that the bacteria were not more negatively affected by low temperatures than phytoplankton.

To freeze above -36.5 degrees C, water requires the presence of an ice nucleus (IN). These can be inert particles or living or dead biological material. As they are the most efficient, inducing freezing at up to -1.8 degrees C, bacteria are the most widely studied biological IN. Here, we show that there is a huge repository of IN in lichens which comprise a large biomass and are able to become airborne. The lichen IN are similar to those we have detected in urban air, exhibiting heat sensitivity but resistance to lysozyme. This suggests many airborne IN are non-bacterial and that eukaryotic IN may be more important to atmospheric processes than previously thought. Copyright. (C) 2009 Royal Meteorological Society

During the PeECE III mesocosm experiment in 2005 we investigated how the virioplankton community responded to increased levels of nutrients (N and P) and CO2. We applied a combination of flow cytometry, Pulsed Field Gel Electrophoresis and degenerate PCR primers to categorize and quantify individual viral populations, and to investigate their temporal dynamics. Species specific and degenerate primers enabled us to identify two specific large dsDNA viruses, EhV and CeV, infecting the haptophytes Emiliania huxleyi and Crysochromulina ericina, respectively. Some of the viral populations detected and enumerated by flow cytometry did not respond to altered CO2-levels, but the abundance of EhV and an unidentified dsDNA virus decreased with increasing CO2 levels. Our results thus indicate that CO2 conditions, or the related change in pH, may affect the marine pelagic food web at the viral level. Our results also demonstrate that in order to unravel ecological problems as how CO2 and nutrient levels affect the relationship between marine algal viruses and their hosts, we need to continue the effort to develop molecular markers used to identify both hosts and viruses.

The predicted rise in anthropogenic CO2 emissions will increase CO2 concentrations and decrease seawater pH in the upper ocean. Recent studies have revealed effects of pCO2 induced changes in seawater chemistry on a variety of marine life forms, in particular calcifying organisms. To test whether the predicted increase in pCO2 will directly or indirectly (via changes in phytoplankton dynamics) affect abundance, activities, and community composition of heterotrophic bacteria during phytoplankton bloom development, we have aerated mesocosms with CO2 to obtain triplicates with three different partial pressures of CO2 (pCO2): 350 μatm (1×CO2), 700 μatm (2×CO2) and 1050 μatm (3×CO2). The development of a phytoplankton bloom was initiated by the addition of nitrate and phosphate. In accordance to an elevated carbon to nitrogen drawdown at increasing pCO2, bacterial production (BPP) of free-living and attached bacteria as well as cell-specific BPP (csBPP) of attached bacteria were related to the C:N ratio of suspended matter. These relationships significantly differed among treatments. However, bacterial abundance and activities were not statistically different among treatments. Solely community structure of free-living bacteria changed with pCO2 whereas that of attached bacteria seemed to be independent of pCO2 but tightly coupled to phytoplankton bloom development. Our findings imply that changes in pCO2, although reflected by changes in community structure of free-living bacteria, do not directly affect bacterial activity. Furthermore, bacterial activity and dynamics of heterotrophic bacteria, especially of attached bacteria, were tightly correlated to phytoplankton development and, hence, may also potentially depend on changes in pCO2.

Feeding the oceans: more carbon is less An experiment in which degradable organic carbon was added to Arctic waters has come up with the counterintuitive finding that in certain conditions, the addition of carbon actually reduces total organic carbon in the ecosystem. The outcome depends on the species interactions within the food web and how they are affected by nutrient supply. When mineral nutrients are limiting, organic carbon accumulates in the system, while in a system limited by organic carbon, marine bacteria are able to outcompete phytoplankton, with the net result a reduction in total organic carbon. This result highlight how descriptions of present and future states of the oceanic carbon cycle depend upon an understanding of the stoichiometric coupling between carbon and growth-limiting mineral nutrients within the marine food web.