Taxonomically and Functionally Distinct Ciliophora Assemblages Inhabiting Baltic Sea Ice.

Ciliophora is a phylum of unicellular eukaryotes that are common and have pivotal roles in aquatic environments. Sea ice is a marine habitat, which is composed of a matrix of solid ice and pockets of saline water in which Ciliophora thrive. Here, we used phylogenetic placement to identify Ciliophora 18S ribosomal RNA reads obtained from wintertime water and sea ice, and assigned functions to the reads based on this taxonomic information. Based on our results, sea-ice Ciliophora assemblages are poorer in taxonomic and functional richness than under-ice water and water-column assemblages. Ciliophora diversity stayed stable throughout the ice-covered season both in sea ice and in water, although the assemblages changed during the course of our sampling. Under-ice water and the water column were distinctly predominated by planktonic orders Choreotrichida and Oligotrichida, which led to significantly lower taxonomic and functional evenness in water than in sea ice. In addition to planktonic Ciliophora, assemblages in sea ice included a set of moderately abundant surface-oriented species. Omnivory (feeding on bacteria and unicellular eukaryotes) was the most common feeding type but was not as predominant in sea ice as in water. Sea ice included cytotrophic (feeding on unicellular eukaryotes), bacterivorous and parasitic Ciliophora in addition to the predominant omnivorous Ciliophora. Potentially mixotrophic Ciliophora predominated the water column and heterotrophic Ciliophora sea ice. Our results highlight sea ice as an environment that creates a set of variable habitats, which may be threatened by the diminishing extent of sea ice due to changing climate.

The first known virus isolates from Antarctic sea ice have complex infection patterns.

Viruses are recognized as important actors in ocean ecology and biogeochemical cycles, but many details are not yet understood. We participated in a winter expedition to the Weddell Sea, Antarctica, to isolate viruses and to measure virus-like particle abundance (flow cytometry) in sea ice. We isolated 59 bacterial strains and the first four Antarctic sea-ice viruses known (PANV1, PANV2, OANV1 and OANV2), which grow in bacterial hosts belonging to the typical sea-ice genera Paraglaciecola and Octadecabacter. The viruses were specific for bacteria at the strain level, although OANV1 was able to infect strains from two different classes. Both PANV1 and PANV2 infected 11/15 isolated Paraglaciecola strains that had almost identical 16S rRNA gene sequences, but the plating efficiencies differed among the strains, whereas OANV1 infected 3/7 Octadecabacter and 1/15 Paraglaciecola strains and OANV2 1/7 Octadecabacter strains. All the phages were cold-active and able to infect their original host at 0°C and 4°C, but not at higher temperatures. The results showed that virus-host interactions can be very complex and that the viral community can also be dynamic in the winter-sea ice.

An active bacterial community linked to high chl-a concentrations in Antarctic winter-pack ice and evidence for the development of an anaerobic sea-ice bacterial community.

Antarctic sea-ice bacterial community composition and dynamics in various developmental stages were investigated during the austral winter in 2013. Thick snow cover likely insulated the ice, leading to high (<4 µg l-1) chlorophyll-a (chl-a) concentrations and consequent bacterial production. Typical sea-ice bacterial genera, for example, Octadecabacter, Polaribacter and Glaciecola, often abundant in spring and summer during the sea-ice algal bloom, predominated in the communities. The variability in bacterial community composition in the different ice types was mainly explained by the chl-a concentrations, suggesting that as in spring and summer sea ice, the sea-ice bacteria and algae may also be coupled during the Antarctic winter. Coupling between the bacterial community and sea-ice algae was further supported by significant correlations between bacterial abundance and production with chl-a. In addition, sulphate-reducing bacteria (for example, Desulforhopalus) together with odour of H2S were observed in thick, apparently anoxic ice, suggesting that the development of the anaerobic bacterial community may occur in sea ice under suitable conditions. In all, the results show that bacterial community in Antarctic sea ice can stay active throughout the winter period and thus possible future warming of sea ice and consequent increase in bacterial production may lead to changes in bacteria-mediated processes in the Antarctic sea-ice zone.

Sea-ice eukaryotes of the Gulf of Finland, Baltic Sea, and evidence for herbivory on weakly shade-adapted ice algae.

To determine community composition and physiological status of early spring sea-ice organisms, we collected sea-ice, slush and under-ice water samples from the Baltic Sea. We combined light microscopy, HPLC pigment analysis and pyrosequencing, and related the biomass and physiological status of sea-ice algae with the protistan community composition in a new way in the area. In terms of biomass, centric diatoms including a distinct Melosira arctica bloom in the upper intermediate section of the fast ice, dinoflagellates, euglenoids and the cyanobacterium Aphanizomenon sp. predominated in the sea-ice sections and unidentified flagellates in the slush. Based on pigment analyses, the ice-algal communities showed no adjusted photosynthetic pigment pools throughout the sea ice, and the bottom-ice communities were not shade-adapted. The sea ice included more characteristic phototrophic taxa (49%) than did slush (18%) and under-ice water (37%). Cercozoans and ciliates were the richest taxon groups, and the differences among the communities arose mainly from the various phagotrophic protistan taxa inhabiting the communities. The presence of pheophytin a coincided with an elevated ciliate biomass and read abundance in the drift ice and with a high Eurytemora affinis read abundance in the pack ice, indicating that ciliates and Eurytemora affinis were grazing on algae.

Solar PAR and UVR modify the community composition and photosynthetic activity of sea ice algae.

The effects of increased photosynthetically active radiation (PAR) and ultraviolet radiation (UVR) on species diversity, biomass and photosynthetic activity were studied in fast ice algal communities. The experimental set-up consisted of nine 1.44 m(2) squares with three treatments: untreated with natural snow cover (UNT), snow-free (PAR + UVR) and snow-free ice covered with a UV screen (PAR). The total algal biomass, dominated by diatoms and dinoflagellates, increased in all treatments during the experiment. However, the smaller biomass growth in the top 10-cm layer of the PAR + UVR treatment compared with the PAR treatment indicated the negative effect of UVR. Scrippsiella complex (mainly Scrippsiella hangoei, Biecheleria baltica and Gymnodinium corollarium) showed UV sensitivity in the top 5-cm layer, whereas Heterocapsa arctica ssp. frigida and green algae showed sensitivity to both PAR and UVR. The photosynthetic activity was highest in the top 5-cm layer of the PAR treatment, where the biomass of the pennate diatom Nitzschia frigida increased, indicating the UV sensitivity of this species. This study shows that UVR is one of the controlling factors of algal communities in Baltic Sea ice, and that increased availability of PAR together with UVR exclusion can cause changes in algal biomass, photosynthetic activity and community composition.

The contribution of mycosporine-like amino acids, chromophoric dissolved organic matter and particles to the UV protection of sea-ice organisms in the Baltic Sea.

The effects of ultraviolet radiation (UVR) on the synthesis of mycosporine-like amino acids (MAAs) in sea-ice communities and on the other UV-absorption properties of sea ice were studied in a three-week long in situ experiment in the Gulf of Finland, Baltic Sea in March 2011. The untreated snow-covered ice and two snow-free ice treatments, one exposed to wavelengths > 400 nm (PAR) and the other to full solar spectrum (PAR + UVR), were analysed for MAAs and absorption coefficients of dissolved (aCDOM) and particulate (ap) fractions, the latter being further divided into non-algal (anap) and algal (aph) components. Our results showed that the diatom and dinoflagellate dominated sea-ice algal community responded to UVR down to 25-30 cm depth by increasing their MAA : chlorophyll-a ratio and by extending the composition of MAA pool from shinorine and palythine to porphyra-334 and an unknown compound with absorption peaks at ca. 335 and 360 nm. MAAs were the dominant absorbing components in algae in the top 10 cm of ice, and their contribution to total absorption became even more pronounced under UVR exposure. In addition to MAAs, the high absorption by chromophoric dissolved organic matter (CDOM) and by deposited atmospheric particles provided UV-protection for sea-ice organisms in the exposed ice. Efficient UV-protection will especially be of importance under the predicted future climate conditions with more frequent snow-free conditions.

Ice formation and growth shape bacterial community structure in Baltic Sea drift ice.

Drift ice, open water and under-ice water bacterial communities covering several developmental stages from open water to thick ice were studied in the northern Baltic Sea. The bacterial communities were assessed with 16S rRNA gene terminal-restriction fragment length polymorphism and cloning, together with bacterial abundance and production measurements. In the early stages, open water and pancake ice were dominated by Alphaproteobacteria and Actinobacteria, which are common bacterial groups in Baltic Sea wintertime surface waters. The pancake ice bacterial communities were similar to the open-water communities, suggesting that the parent water determines the sea-ice bacterial community in the early stages of sea-ice formation. In consolidated young and thick ice, the bacterial communities were significantly different from water bacterial communities as well as from each other, indicating community development in Baltic Sea drift ice along with ice-type changes. The thick ice was dominated by typical sea-ice genera from classes Flavobacteria and Gammaproteobacteria, similar to those in polar sea-ice bacterial communities. Since the thick ice bacterial community was remarkably different from that of the parent seawater, results indicate that thick ice bacterial communities were recruited from the rarer members of the seawater bacterial community.

Rhinomonas nottbecki n. sp. (cryptomonadales) and molecular phylogeny of the family Pyrenomonadaceae.

The cryptomonad Rhinomonas nottbecki n. sp., isolated from the Baltic Sea, is described from live and fixed cells studied by light, scanning, and transmission electron microscopy together with sequences of the partial nucleus- and nucleomorph-encoded 18S rRNA genes as well as the nucleus-encoded ITS1, 5.8S, ITS2, and the 5'-end of the 28S rRNA gene regions. The sequence analyses include comparison with 43 strains from the family Pyrenomonadaceae. Rhinomonas nottbecki cells are dorsoventrally flattened, obloid in shape; 10.0-17.2 µm long, 5.5-8.1 µm thick, and 4.4-8.8 µm wide. The inner periplast has roughly hexagonal plates. Rhinomonas nottbecki cells resemble those of Rhinomonas reticulata, but the nucleomorph 18S rRNA gene of R. nottbecki differs by 2% from that of R. reticulata, while the ITS region by 11%. The intraspecific variability in the ITS region of R. nottbecki is 5%. In addition, the predicted ITS2 secondary structures are different in R. nottbecki and R. reticulata. The family Pyrenomonadaceae includes three clades: Clade A, Clade B, and Clade C. All Rhinomonas sequences branched within the Clade C, while the genus Rhodomonas is paraphyletic. The analyses suggest that the genus Storeatula is an alternating morphotype of the genera Rhinomonas and Rhodomonas and that the family Pyrenomonadaceae includes some species that were described multiple times, as well as novel species.

Isolation and characterization of phage-host systems from the Baltic Sea ice.

In search for sea ice bacteria and their phages from the Baltic Sea ice, two ice samples were collected from land-fast ice in a south-west Finland coastal site in February and March 2011. Bacteria were isolated from the melted sea ice samples and phages were screened from the same samples for 43 purified isolates. Plaque-producing phages were found for 15 bacterial isolates at 3 °C. Ten phage isolates were successfully plaque purified and eight of them were chosen for particle purification to analyze their morphology and structural proteins. Phage 1/32 infecting an isolate affiliated to phylum Bacteroidetes (Flavobacterium sp.) is a siphovirus and six phages infecting isolates affiliated to γ-Proteobacteria (Shewanella sp.) hosts were myoviruses. Cross titrations between the hosts showed that all studied phages are host specific. Phage solutions, host growth and phage infection were tested in different temperatures revealing phage temperature tolerance up to 45 °C, whereas phage infection was in most of the cases retarded above 15 °C. This study is the first to report isolation and cultivation of ice bacteria and cold-active phages from the Baltic Sea ice.

Role of sea-ice biota in nutrient and organic material cycles in the northern Baltic Sea.

This paper compiles biological and chemical sea-ice data from three areas of the Baltic Sea: the Bothnian Bay (Hailuoto, Finland), the Bothnian Sea (Norrby, Sweden), and the Gulf of Finland (Tvärminne, Finland). The data consist mainly of field measurements and experiments conducted during the BIREME project from 2003 to 2006, supplemented with relevant published data. Our main focus was to analyze whether the biological activity in Baltic Sea sea-ice shows clear regional variability. Sea-ice in the Bothnian Bay has low chlorophyll a concentrations, and the bacterial turnover rates are low. However, we have sampled mainly land-fast level first-year sea-ice and apparently missed the most active biological system, which may reside in deformed ice (such as ice ridges). Our limited data set shows high concentrations of algae in keel blocks and keel block interstitial water under the consolidated layer of the pressure ridges in the northernmost part of the Baltic Sea. In land-fast level sea-ice in the Bothnian Sea and the Gulf of Finland, the lowermost layer appears to be the center of biological activity, though elevated biomasses can also be found occasionally in the top and interior parts of the ice. Ice algae are light limited during periods of snow cover, and phosphate is generally the limiting nutrient for ice bottom algae. Bacterial growth is evidently controlled by the production of labile dissolved organic matter by algae because low growth rates were recorded in the Bothnian Bay with high concentrations of allochthonous dissolved organic matter. Bacterial communities in the Bothnian Sea and the Gulf of Finland show high turnover rates, and activities comparable with those of open water communities during plankton blooms, which implies that sea-ice bacterial communities have high capacity to process matter during the winter period.