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Special issue Polar Microbes.-- 22 pages, 4 figures, 5 tables, supplementary material https://doi.org/10.3390/microorganisms9020317.-- Data Availability Statement: All data is reported in the present article.-- This research is part of POLARCSIC activities, The ocean surface microlayer (SML), with physicochemical characteristics different from those of subsurface waters (SSW), results in dense and active viral and microbial communities that may favor virus–host interactions. Conversely, wind speed and/or UV radiation could adversely affect virus infection. Furthermore, in polar regions, organic and inorganic nutrient inputs from melting ice may increase microbial activity in the SML. Since the role of viruses in the microbial food web of the SML is poorly understood in polar oceans, we aimed to study the impact of viruses on prokaryotic communities in the SML and in the SSW in Arctic and Antarctic waters. We hypothesized that a higher viral activity in the SML than in the SSW in both polar systems would be observed. We measured viral and prokaryote abundances, virus-mediated mortality on prokaryotes, heterotrophic and phototrophic nanoflagellate abundance, and environmental factors. In both polar zones, we found small differences in environmental factors between the SML and the SSW. In contrast, despite the adverse effect of wind, viral and prokaryote abundances and virus-mediated mortality on prokaryotes were higher in the SML than in the SSW. As a consequence, the higher carbon flux released by lysed cells in the SML than in the SSW would increase the pool of dissolved organic carbon (DOC) and be rapidly used by other prokaryotes to grow (the viral shunt). Thus, our results suggest that viral activity greatly contributes to the functioning of the microbial food web in the SML, which could influence the biogeochemical cycles of the water column, This research was part of the ATOS project, funded as part of the Spanish contribution to the International Polar Year (IPY) by the Spanish Ministry of Science and Innovation (POL200600550/CTM). J.A.B.’s work was supported by a PhD fellowship from the Spanish Ministerio de Ciencia e Innovación (FPU grant), With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)

Ocean acidification and warming are two main consequences of climate change that can directly affect biological and ecosystem processes in marine habitats. The Arctic Ocean is the region of the world experiencing climate change at the steepest rate compared with other latitudes. Since marine planktonic microorganisms play a key role in the biogeochemical cycles in the ocean it is crucial to simultaneously evaluate the effect of warming and increasing CO2 on marine microbial communities. In 20 L experimental microcosms filled with water from a high-Arctic fjord (Svalbard), we examined changes in phototrophic and heterotrophic microbial abundances and processes [bacterial production (BP) and mortality], and viral activity (lytic and lysogenic) in relation to warming and elevated CO2. The summer microbial plankton community living at 1.4°C in situ temperature, was exposed to increased CO2 concentrations (135–2,318 μatm) in three controlled temperature treatments (1, 6, and 10°C) at the UNIS installations in Longyearbyen (Svalbard), in summer 2010. Results showed that chlorophyll a concentration decreased at increasing temperatures, while BP significantly increased with pCO2 at 6 and 10°C. Lytic viral production was not affected by changes in pCO2 and temperature, while lysogeny increased significantly at increasing levels of pCO2, especially at 10°C (R2 = 0.858, p = 0.02). Moreover, protistan grazing rates showed a positive interaction between pCO2 and temperature. The averaged percentage of bacteria grazed per day was higher (19.56 ± 2.77% d-1) than the averaged percentage of lysed bacteria by virus (7.18 ± 1.50% d-1) for all treatments. Furthermore, the relationship among microbial abundances and processes showed that BP was significantly related to phototrophic pico/nanoflagellate abundance in the 1°C and the 6°C treatments, and BP triggered viral activity, mainly lysogeny at 6 and 10°C, while bacterial mortality rates was significantly related to bacterial abundances at 6°C. Consequently, our experimental results suggested that future increases in water temperature and pCO2 in Arctic waters will produce a decrease of phytoplankton biomass, enhancement of BP and changes in the carbon fluxes within the microbial food web. All these heterotrophic processes will contribute to weakening the CO2 sink capacity of the Arctic plankton community.

The microbial response to environmental changes in coastal waters of the eastern Cantabrian Sea was explored for four years by analysing a broad set of environmental variables along with bacterial community metabolism and composition. A recurrent seasonal cycle emerged, consisting of two stable periods, characterized by low bacterial metabolic activity (winter) from October to March, and high bacterial metabolic activity (summer) from May to August. These two contrasting periods were linked by short transition periods in April (TA ) and September (TS ). The phylogenetic groups Alphaproteobacteria and Bacteroidetes were dominant during winter and summer respectively, and their recurrent alternation was mainly driven by the bloom of eukaryotic phytoplankton before TA and the bloom of prokaryotic phytoplankton before TS . Bacterial growth efficiency remained high and stable during the winter and summer periods but dropped during the two short transition periods. Our results suggest that bacterial growth efficiency should be considered a very resilient property that reflects different stages in the adaptation of the bacterial community composition to the environmental changes occurring throughout the seasonal cycle in this coastal ecosystem., SI

12 pages, 4 figures, 3 tables, supplementary material http://journal.frontiersin.org/article/10.3389/fmicb.2017.00241/full#supplementary-material, During the Austral summer 2009 we studied three areas surrounding the Antarctic Peninsula: the Bellingshausen Sea, the Bransfield Strait and the Weddell Sea. We aimed to investigate, whether viruses or protists were the main agents inducing prokaryotic mortality rates, and the sensitivity to temperature of prokaryotic heterotrophic production and mortality based on the activation energy (Ea) for each process. Seawater samples were taken at seven depths (0.1–100 m) to quantify viruses, prokaryotes and protists abundances, and heterotrophic prokaryotic production (PHP). Viral lytic production, lysogeny, and mortality rates of prokaryotes due to viruses and protists were estimated at surface (0.1–1 m) and at the Deep Fluorescence Maximum (DFM, 12–55 m) at eight representative stations of the three areas. The average viral lytic production ranged from 1.0 ± 0.3 × 107 viruses ml−1 d−1 in the Bellingshausen Sea to1.3 ± 0.7 × 107 viruses ml−1 d−1 in the Bransfield Strait, while lysogeny, when detectable, recorded the lowest value in the Bellingshausen Sea (0.05 ± 0.05 × 107 viruses ml−1 d−1) and the highest in the Weddell Sea (4.3 ± 3.5 × 107 viruses ml−1 d−1). Average mortality rates due to viruses ranged from 9.7 ± 6.1 × 104 cells ml−1 d−1 in the Weddell Sea to 14.3 ± 4.0 × 104 cells ml−1 d−1 in the Bellingshausen Sea, and were higher than averaged grazing rates in the Weddell Sea (5.9 ± 1.1 × 104 cells ml−1 d−1) and in the Bellingshausen Sea (6.8 ± 0.9 × 104 cells ml−1 d−1). The highest impact on prokaryotes by viruses and main differences between viral and protists activities were observed in surface samples: 17.8 ± 6.8 × 104 cells ml−1 d−1 and 6.5 ± 3.9 × 104 cells ml−1 d−1 in the Weddell Sea; 22.1 ± 9.6 × 104 cells ml−1 d−1 and 11.6 ± 1.4 × 104 cells ml−1 d−1 in the Bransfield Strait; and 16.1 ± 5.7 × 104 cells ml−1 d−1 and 7.9 ± 2.6 × 104 cells ml−1 d−1 in the Bellingshausen Sea, respectively. Furthermore, the rate of lysed cells and PHP showed higher sensitivity to temperature than grazing rates by protists. We conclude that viruses were more important mortality agents than protists mainly in surface waters and that viral activity has a higher sensitivity to temperature than grazing rates. This suggests a reduction of the carbon transferred through the microbial food-web that could have implications in the biogeochemical cycles in a future warmer ocean scenario, This study was supported by the following projects: ATOS (POL2006-00550/CTM, P.I.: CD.) funded by the Spanish Ministerio de Ciencia e Innovación, YC work was supported by a Ph.D. fellowship from the MINECO (FPI grant)

Pernice, Massimo ... et. al.-- 11 pages, 7 figures, 3 tables, The dark ocean is one of the largest biomes on Earth, with critical roles in organic matter remineralization and global carbon sequestration. Despite its recognized importance, little is known about some key microbial players, such as the community of heterotrophic protists (HP), which are likely the main consumers of prokaryotic biomass. To investigate this microbial component at a global scale, we determined their abundance and biomass in deepwater column samples from the Malaspina 2010 circumnavigation using a combination of epifluorescence microscopy and flow cytometry. HP were ubiquitously found at all depths investigated down to 4000 m. HP abundances decreased with depth, from an average of 72±19 cells ml-1 in mesopelagic waters down to 11±1 cells ml-1 in bathypelagic waters, whereas their total biomass decreased from 280±46 to 50±14 pg C ml-1. The parameters that better explained the variance of HP abundance were depth and prokaryote abundance, and to lesser extent oxygen concentration. The generally good correlation with prokaryotic abundance suggested active grazing of HP on prokaryotes. On a finer scale, the prokaryote:HP abundance ratio varied at a regional scale, and sites with the highest ratios exhibited a larger contribution of fungi molecular signal. Our study is a step forward towards determining the relationship between HP and their environment, unveiling their importance as players in the dark ocean's microbial food web. © 2015 International Society for Microbial Ecology All rights reserved, This study was supported by the Spanish Ministry of Science and Innovation through project Consolider-Ingenio Malaspina 2010 (CSD2008-00077) (to CMD) and FLAME (CGL2010-16304) (to RM)

© 2014 Mayol, Jiménez, Herndl, Duarte and Arrieta. Airborne transport of microbes may play a central role in microbial dispersal, the maintenance of diversity in aquatic systems and in meteorological processes such as cloud formation. Yet, there is almost no information about the abundance and fate of microbes over the oceans, which cover >70% of the Earth's surface and are the likely source and final destination of a large fraction of airborne microbes. We measured the abundance of microbes in the lower atmosphere over a transect covering 17° of latitude in the North Atlantic Ocean and derived estimates of air-sea exchange of microorganisms from meteorological data. The estimated load of microorganisms in the atmospheric boundary layer ranged between 6×104 and 1.6×107 microbes per m2 of ocean, indicating a very dynamic air-sea exchange with millions of microbes leaving and entering the ocean per m2 every day. Our results show that about 10% of the microbes detected in the boundary layer were still airborne 4 days later and that they could travel up to 11,000 km before they entered the ocean again. The size of the microbial pool hovering over the North Atlantic indicates that it could play a central role in the maintenance of microbial diversity in the surface ocean and contribute significantly to atmospheric processes., This is a contribution to the Malaspina Expedition 2010, funded by the INGENIO 2010 CONSOLIDER program (ref. CDS2008-00077) of the Spanish Ministry of Economy and Competitiveness, and projects from the Austrian Science Fund (FWF, I486-B09 and P23234-B11) and project MEDEA from the European Research Council under the European Community's Seventh Framework Programme (FP7/2007-2013, ERC grant agreement No. 268595). Eva Mayol and María A. Jiménez acknowledge the “Junta para la Ampliación de Estudios” program (JAE-predoc and JAE-doc contracts, respectively) from CSIC, supplied by the European Social Fund. Jesús M. Arrieta was supported by a “Ramón y Cajal” fellowship by the former Ministry of Science and Innovation of the Spanish Government

Bacterial endophytes are crucial for the survival of many terrestrial plants, but little is known about the presence and importance of bacterial endophytes of marine plants. We conducted a survey of the endophytic bacterial community of the long-living Mediterranean marine angiosperm Posidonia oceanica in surface-sterilized tissues (roots, rhizomes, and leaves) by Denaturing Gradient Gel Electrophoresis (DGGE). A total of 26 Posidonia oceanica meadows around the Balearic Islands were sampled, and the band patterns obtained for each meadow were compared for the three sampled tissues. Endophytic bacterial sequences were detected in most of the samples analyzed. A total of 34 OTUs (Operational Taxonomic Units) were detected. The main OTUs of endophytic bacteria present in P. oceanica tissues belonged primarily to Proteobacteria (¿, ¿, and ¿ subclasses) and Bacteroidetes. The OTUs found in roots significantly differed from those of rhizomes and leaves. Moreover, some OTUs were found to be associated to each type of tissue. Bipartite network analysis revealed differences in the bacterial endophyte communities present on different islands. The results of this study provide a pioneering step toward the characterization of the endophytic bacterial community associated with tissues of a marine angiosperm and reveal the presence of bacterial endophytes that differed among locations and tissue types.

Clones from the same marine bacterioplankton community were sequenced, 100 clones based on DNA (16S rRNA genes) and 100 clones based on RNA (16S rRNA). This bacterioplankton community was dominated by alpha-Proteobacteria in terms of repetitive DNA clones (52%), but gamma-Proteobacteria dominated in terms of repetitive RNA clones (44%). The combined analysis led to a characterization of phylotypes otherwise uncharacterized if only the DNA or RNA libraries would have been analyzed alone. Of the DNA clones, 25.5% were found only in this library and no close relatives were detected in the RNA library. For clones from the RNA library, 21.5% of RNA clones did not indicate close relatives in the DNA library. Based on the comparisons between DNA and RNA libraries, our data indicate that the characterization of the bacterial community based on RNA has the potential to characterize distinct phylotypes from the marine environment, which remain undetected on the DNA level.

T-RFLP clone characterization screening was optimized for a fast and basepair-accurate characterization of clones from marine Archaea collected from the Eastern Mediterranean Sea. Because of the high sensitivity of T-RFLP fingerprinting, a protocol was developed where 10 initial PCR cycles gave detectable terminal fragments from clones. Additionally, forward and reverse primers for PCR were individually labeled and detected simultaneously to assess the suitability of the forward and reverse fragments for T-RFLP screening. Based on independent restriction digests with the tetrameric restriction enzymes HhaI, RsaI and HaeIII to characterize the 49 archaeal clones in our library, the clones were grouped into 13 . T-RFLP operational taxonomic units OTUs . Reverse fragments generally gave less heterogeneous fragments in size. The . accuracy of T-RFLP screening was evaluated by sequencing representative clones. Closely related clones f 97% similarity could only be resolved with multiple restriction digests where forward and reverse fragments were included in the analysis. All fragments from the clone library were detected in the T-RFLP fingerprint from the complex archaeal community. We found representatives of marine group I, II and III Archaea. Thus, the recently discovered low abundant marine group III Archaea could be clearly differentiated from the other clones in our library and comprised a considerable fraction of the . clone library f 12% . Therefore, our T-RFLP screening approach proved successful in characterizing novel archaeal sequences from the marine environment. q 2001 Published by Elsevier Science B.V.

Two kinds of particles were created in the laboratory in rolling tanks to simulate natural macroaggregates. Laboratory-made particles type 1 were formed under sterile conditions from a mixture of cells and extracellular products of four diatom species in artificial seawater, and inoculated with a marine microbial assemblage. Laboratory-made particles type 2 were created directly from natural seawater. These particles were characterised in terms of maximum length, volume and abundance. Chemical composition (carbohydrates, amino acids and total organic carbon) and bacterial and flagellate abundances were measured in the particles and ambient water. We found that both kinds of laboratory-made particles were similar in terms of size, chemical composition and microbial abundance. Moreover, they resembled natural marine aggregates in size and volume. However, laboratory-made particles showed higher concentrations of carbohydrates, amino acids and total organic carbon as well as higher microbial abundance when compared to natural macroaggregates. This difference can be explained by the sampling method, since natural aggregates are frequently collected in the sea with syringes including ambient water, and consequently diluted, whereas in this study the laboratory-made particles were carefully collected without ambient water. Thus, both kinds of laboratory-made particles might be a good alternative for the analysis of microbial processes in marine macroaggregates. Advantages and disadvantages of these two types of laboratory-made particles are discussed.