Microbial community development on model particles in the deep sulfidic waters of the Black Sea

Microorganisms attached to particles have been shown to be different from free-living microbes and to display diverse metabolic activities. However, little is known about the ecotypes associated with particles and their substrate preference in anoxic marine waters. Here, we investigate the microbial community colonizing particles in the anoxic and sulfide-rich waters of the Black Sea. We incubated beads coated with different substrates in situ at 1000 and 2000 m depth. After 6 h, the particle-attached microbes were dominated by Gamma- and Alpha-proteobacteria, and groups related to the phyla Latescibacteria, Bacteroidetes, Planctomycetes and Firmicutes, with substantial variation across the bead types, indicating that the attaching communities were selected by the substrate. Further laboratory incubations for 7 days suggested the presence of a community of highly specialized taxa. After incubation for 35 days, the microbial composition across all beads and depths was similar and primarily composed of putative sulfur cycling microbes. In addition to the major shared microbial groups, subdominant taxa on chitin and protein-coated beads were detected pointing to specialized microbial degraders. These results highlight the role of particles as sites for attachment and biofilm formation, while the composition of organic matter defined a secondary part of the microbial community.     

Diversity and distribution of Planctomycetes and related bacteria in the suboxic zone of the Black Sea

Samples from six depths of the Black Sea's suboxic zone were analyzed for 16S rRNA gene sequence information. A gradient in phylotype diversity was found. The distributions of known anaerobic ammonium oxidation (anammox) bacteria, many unknown Planctomycetes, and other phylotypes were examined in relation to the local nutrient and redox conditions.     

Physiological,chemotaxonomic and genomic characterization of two novel piezotolerant bacteria of the family Marinifilaceae isolated from sulfidic waters of the Black Sea

Diversity analyses of microbial enrichments obtained from deep sulfidic water (2000 m) collected from the Black Sea indicated the presence of eleven novel putative lineages of bacteria affiliated to the family Marinifilaceae of the phylum Bacteroidetes. Pure cultures were obtained for four strains (i.e. M1P^T, M3P, A4^T and 44) of this family, which could be grouped into two different clades based on their 16S rRNA gene sequences. All four strains were Gram-negative, rod-shaped and facultative anaerobic bacteria. The genomes of all strains were sequenced and physiological analyses were performed. All strains utilized a wide range of carbon sources, which was supported by the presence of the pathways involved in carbon utilization encoded by their genomes. The strains were able to grow at elevated hydrostatic pressure (up to 50 MPa), which coincided with increased production of unsaturated and branched fatty acids, and a decrease in hydroxy fatty acids. Intact polar lipid analysis of all four strains showed the production of ornithine lipids, phosphatidylethanolamines and capnine lipids as major intact polar lipids (IPLs). Genes involved in hopanoid biosynthesis were also identified. However, bacteriohopanepolyols (BHPs) were not detected in the strains. Based on distinct physiological, chemotaxonomic, genotypic and phylogenetic differences compared to other members of the genera Ancylomarina and Labilibaculum, it was concluded that strains M1P^T and A4^T represented two novel species for which the names Ancylomarina euxinus sp. nov. and Labilibaculum euxinus sp. nov., respectively, are proposed.     

Putative ammonia-oxidizing Crenarchaeota in suboxic waters of the Black Sea: a basin-wide ecological study using 16S ribosomal and functional genes and membrane lipids

Within the upper 400 m at western, central and eastern stations in the world's largest stratified basin, the Black Sea, we studied the qualitative and quantitative distribution of putative nitrifying Archaea based on their genetic markers (16S rDNA, amoA encoding for the alpha-subunit of archaeal ammonia monooxygenase), and crenarchaeol, the specific glycerol diphytanyl glycerol tetraether of pelagic Crenarchaeota within the Group I.1a. Marine Crenarchaeota were the most abundant Archaea (up to 98% of the total archaeal 16S rDNA copies) in the suboxic layers with oxygen levels as low as 1 microM including layers where previously anammox bacteria were described. Different marine crenarchaeotal phylotypes (both 16S rDNA and amoA) were found at the upper part of the suboxic zone as compared with the base of the suboxic zone and the upper 15-30 m of the anoxic waters with prevailing sulfide concentrations of up to 30 microM. Crenarchaeol concentrations were higher in the sulfidic chemocline as compared with the suboxic zone. These results indicate an abundance of putative nitrifying Archaea at very low oxygen levels within the Black Sea and might form an important source of nitrite for the anammox reaction.     

Metabolic strategies of free-living and aggregate-associated bacterial communities inferred from biologic and chemical profiles in the Black Sea suboxic zone

The Black Sea is a permanently anoxic basin with a well-defined redox gradient. We combine environmental 16S rRNA gene data from clone libraries, terminal restriction fragment length polymorphisms, and V6 hypervariable region pyrosequences to provide the most detailed bacterial survey to date. Furthermore, this data set is informed by comprehensive geochemical data; using this combination of information, we put forward testable hypotheses regarding possible metabolisms of uncultured bacteria from the Black Sea's suboxic zone (microaerophily, nitrate reduction, manganese cycling, and oxidation of methane, ammonium, and sulfide). Dominant bacteria in the upper suboxic zone included members of the SAR11, SAR324, and Microthrix groups and in the deep suboxic zone included members of BS-GSO-2, Marine Group A, and SUP05. A particulate fraction (30?μm filter) was used to distinguish between free-living and aggregate-attached communities in the suboxic zone. The particulate fraction contained greater diversity of V6 tag sequences than the bulk water samples. Lentisphaera, Epsilonproteobacteria, WS3, Planctomycetes, and Deltaproteobacteria were enriched in the particulate fraction, whereas SAR11 relatives dominated the free-living fraction. On the basis of the bacterial assemblages and simple modeling, we find that in suboxic waters, the interior of sinking aggregates potentially support manganese reduction, sulfate reduction, and sulfur oxidation.     

Marine biological invasions in waters of the port of Novorossiysk in the Black Sea

The results of the monitoring of ballast waters of commercial vessels and research on the ecosystem biodiversity of the Novorossiysk Port are reported. Data on structure of immigrant fauna, its abundance, survival and origin of species were obtained. It has been established that the risk of introduction of Mediterranean species into the Novorossiysk Bay is the most probable. The basic “risk groups” are copepods and polychaetes. It has been hypothesized, that ship ballast water can become a primary factor of the “mediterranization” of the copepod fauna (Copepoda) in the northeast of the Black Sea. During the period of research in the Novorossiysk Bay, 36 species of Mediterranean copepods were recorded. A massive number of the cyclopoid copepod Oithona brevicornis Giesbr., 1891, a new species for the Black Sea, was recorded in samples collected in the autumn of 2005 and 2006. The status of the invader was attributed to the polychaete of the Streblospio genus. The necessity of ship ballast water control in Russia and the development of methodology, methods, and regulations for the prevention of the intrusion of pathogenic and potentially dangerous organisms by marine traffic were corroborated.     

Protist diversity and distribution: some basic considerations

This essay discusses protist species number and geographic distribution, both heavily influenced by undersampling and human introductions. The features of the ubiquity model and the moderate endemicity model are compared. I recognize five main flaws of the ubiquity model, viz., the ignorance of the extraordinary possibilities protists have to speciate due to their short generation time and the likelihood that many persisted over geological time scales; that all protist species have high abundances; that their small size is a main reason for global distribution; the ignorance of human introductions; and the rejection of literature evidence on the occurrence of flagship species with restricted distribution in a wide variety of protists. Thus, the data available support the moderate endemicity model which proposes about 300,000 extant, free-living protist species, of which one third might have a restricted distribution, i.e., is not cosmopolitan in spite of suitable habitats. To sum up, the distribution of protists, flowering plants, and larger animals has much in common, but protists usually have wider ranges and thus a higher proportion of cosmopolites. Future research should reconcile morphologic, genetic, and ecological species concepts because this is crucial for determining the number of protist species. Further, greatly intensified research is required on morphospecies in heterotrophic protists because their diversity has never been investigated in large areas of the earth. Special Issue: Protist diversity and geographic distribution. Guest editor: W. Foissner.     

Analysis of nitrite reductase (nirK and nirS) genes and cultivation reveal depauperate community of denitrifying bacteria in the Black Sea suboxic zone

Chemical profiles of the Black Sea suboxic zone show a distribution of nitrogen species which is traditionally associated with denitrification, i.e. a secondary nitrite maximum associated with nitrate depletion and a N(2) gas peak. To better understand the distribution and diversity of the denitrifier community in the Black Sea suboxic zone, we combined a cultivation approach with cloning and sequencing of PCR-amplified nitrite reductase (nirS and nirK) genes. The Black Sea suboxic zone appears to harbour a homogeneous community of denitrifiers. For nirK, over 94% of the sequences fell into only three distinct phylogenetic clusters, and for nirS, a single closely related sequence type accounted for 91% of the sequences retrieved. Both nirS and nirK genes showed a dramatic shift in community composition at the bottom of the suboxic zone, but overall, nirK-based community composition showed much greater variation across depths compared with the highly uniform distribution of nirS sequences throughout the suboxic zone. The dominant nirK and nirS sequences differed at the amino acid level by at least 17% and 8%, respectively, from their nearest database matches. Denitrifying isolates recovered from the suboxic zone shared 97% 16S rRNA gene sequence similarity with Marinobacter maritimus. Analysis of the recently discovered nirS gene from the anammox bacterium Candidatus'Kuenenia stuttgartiensis' revealed that mismatches with commonly used primers may have prevented the previous detection of this divergent sequence.     

Ichthyoplankton of the coastal waters of northeastern shelf of the Black Sea and the Kerch Strait

We study the state of summer ichthyoplankton and dynamics of its taxonomic composition and abundance near large harbor cities (Novorossiysk and Tuapse), as well as in the navigation areas and near transshipment harbor facilities (Taman?? Prichernomor??e and Kerch Strait). We show that temporal-spatial changes in the taxonomic composition and abundance of ichthyoplankton are determined by fluctuations of water temperature, pressure of predatory comb jelly Mnemiopsis leydyi and level of water pollution. High mortality rate and low abundance of fish eggs and larvae in the polluted areas of Novorossiysk, Tuapse, and Kerch Strait indicate unfavorable conditions for the embryonic and postembryonic development of fish in these areas. As judged by the state of ichthyoplankton the most favorable conditions are in the open part of the Novorossiysk Bay and Taman?? Prichernomor??e. Starting from 2002, the spawning populations of pelagophylic fish in the Novorossiysk Bay are gradually recovering to the level of 1980.     

Heterotrophic bacteria,zooflagellates, and ciliates in coastal waters of northeastern Black Sea

According to the parameters of density and biomass of heterotrophic bacterioplankton, waters in the ports of Novorossiysk and Tuapse were eutrophic-hypereutrophic and in the resort cities of Gelendzhik and Anapa they were hypereutrophic. The abundance of heterotrophic bacteria reached maximum values of 12.7–14.2 million cells/mL during the period of abnormally high water temperatures (August 2010) in recreational zones. Chainlike and filamentous forms (57–65%) of bacteria, which were typical for the strong coastal pollution of waters, prevailed. The abundance of zooflagellates (kinetoplastids) in semi-isolated water areas (port of Novorossiysk and Gelendzhik Bay) reached the level of highly eutrophic waters, 6.2–9.7 million ind/m³. However, the biomass of naked ciliates was 1.5 times lower than their maximum values earlier registered in coastal waters of the northeastern shelf. The peak of abundance of alien tintinnids of the genera Eutintinnus, Tintinnopsis, and Amphorellopsis, which were introduced via ballast waters of ships, was recorded in Novorossiysk Bay. The ratio of titinnids to the total abundance of ciliates increased 5 times and reached 25–40%. Protozoans developed poorly in the oil-polluted port of Tuapse and the open Anapa Bay.     

Microbiological processes at the interface of aerobic and anaerobic waters in the deep-water zone of the Black Sea

Chemical and key microbiological processes (assimilation of carbon dioxide, oxidation and formation of methane, and sulfate reduction) occurring at the boundary between the aerobic-anaerobic interface in the deep-water zone of the Black Sea were investigated. Measurements were taken at depths from 90 to 300 m at intervals of 5-10 m. The integral rate of the dark assimilation of carbon dioxide varied from 120 to 207 mg C/(m2 day) with a maximum at the boundary of cyclonic currents. The organic matter (OM) formed from methane comprised less than 5% of the OM formed from carbon dioxide. A comparison between the rates of methane oxidation and methane production suggests that methane that is oxidized at depths from 100 to 300 m was formed in deeper water horizons. The maximum rate of sulfate reduction (1230 mg S/(m2 day)) was observed in the western halistatic region, and the minimum rate (490 mg S/(m2 day)), in the eastern halistatic region. The average rate of hydrogen sulfide production measured at three deep-sea stations amounted to 755 mg S/(m2 day), or 276 g S/(m2 year).     

Structure and seasonal trophodynamics of picophytoplankton in sevastopol bay and adjacent waters (the Black Sea)

Abundance and seasonal trophodynamics (specific growth rate, daily production, and grazing mortality) of the major picophytoplankton components, Synechococcus cyanobacteria (Syn) and picoeukaryotes (Pico-E), were studied at three stations in Sevastopol Bay and adjacent coastal waters (the Black Sea) in 2014 by flow cytometry and the dilution method. Pico-E abundance was shown to increase along the nutrient and pollution gradient from the coastal waters outside the bay (annual average of 7.3 ± 5.4 × 10³ cells mL^–1) to the eastern corner of the bay (28.7 ± 11.4 × 10³ cells mL^–1), while no relation was found between the water pollution status and Syn abundance (9.9 ± 8.7 × 10³ cells mL^–1; at all the stations, n = 27). Matter flows through the communities (daily production for Syn and Pico-E 0^–16.6 and 0^–19.3 µg C L^–1 day^–1, respectively; grazing mortality for Syn and PicoE 0–3.6 and 0–21.2 µg C L^–1 day^–1, respectively) were comparable to or even exceeded their biomass stocks (>0.05–6.8 and 0.9–26.5 µg C L^–1 for Syn and PicoE, respectively), indicating high biomass turnover rates. The highest flow-to-stock ratio (up to 6 for Syn) and a significant imbalance between daily production (P) and grazing mortality (G) were observed in the most polluted and eutrophic waters of the bay in spring (Pico-E: P/G < 1) and late summer (Syn: P/G > 1). Black River inflow to the bay was hypothesized to be among the mechanisms maintaining this pronounced and long-term imbalance in the open system without any negative consequences for the picophytoplankton assemblages.     

Spatial distribution and diversity of commercial demersal fish species in the shelf waters of the Caspian Sea

The Caspian Sea (CS) is a unique ecosystem known for its several fish species, especially sturgeons. To exhaustively manage the fish stocks of this ecosystem, detailed knowledge of species composition, abundance, distribution, and the habitat traits of the living organisms is necessary. This study analyzed the diversity, spatial, and seasonal distribution of commercial demersal fish species, and examined the relationships between community structure and environmental variability in Iranian shelf waters of the CS. For this purpose, seasonally fish sampling took place between 2009 and 2011 with a bottom trawl. Among 11 fish species captured, Chelon spp., Rutilus kutum, and Vimba vimba showed the highest abundances (i.e. 88.49%, 10.67%, and 0.69%, respectively). Univariate and multivariate analyses showed differences in commercial fish assemblages (abundance and species richness) according to seasons and regions. Distance-based Linear Model (DisTLM) showed that eight environmental variables display significant linear relationships with the fauna resemblance matrix (p < .05). Based on AIC criteria, the combination of silt&clay, TOM, longitude, depth, bottom, and surface temperature used to build the parsimonious DisTLM model explain 67.03% of the total variability. The results revealed a biogeographical and temporal gradient from the west to the east and summer to winter, in terms of commercial demersal fish assemblages and species diversity, as a consequence of different geomorphological, bottom substratum conditions and benthic communities.     

Protist diversity in a permanently ice-covered Antarctic Lake during the polar night transition

The McMurdo Dry Valleys of Antarctica harbor numerous permanently ice-covered lakes, which provide a year-round oasis for microbial life. Microbial eukaryotes in these lakes occupy a variety of trophic levels within the simple aquatic food web ranging from primary producers to tertiary predators. Here, we report the first molecular study to describe the vertical distribution of the eukaryotic community residing in the photic zone of the east lobe (ELB) and west lobe (WLB) of the chemically stratified Lake Bonney. The 18S ribosomal RNA (rRNA) libraries revealed vertically stratified populations dominated by photosynthetic protists, with a cryptophyte dominating shallow populations (ELB–6 m; WLB–10 m), a haptophyte occupying mid-depths (both lobes 13 m) and chlorophytes residing in the deepest layers (ELB–18 and 20 m; WLB–15 and 20 m) of the photic zone. A previously undetected stramenopile occurred throughout the water column of both lobes. Temporal variation in the eukaryotic populations was examined during the transition from Antarctic summer (24-h sunlight) to polar night (complete dark). Protist diversity was similar between the two lobes of Lake Bonney due to exchange between the photic zones of the two basins via a narrow bedrock sill. However, vertical and temporal variation in protist distribution occurred, indicating the influence of the unique water chemistry on the biology of the two dry valley watersheds.     

16S rRNA-based bacterial diversity in the organic-rich sediments underlying oxygen-deficient waters of the eastern Arabian Sea

The eastern Arabian Sea has a unique and permanent oxygen minimum zone (OMZ) that extends along the western continental margin of India. The sediment below this region is rich in organic matter. This study describes the bacterial community structure and diversity in OMZ sediments of the eastern Arabian Sea (AS) through 16S rRNA clone library analysis. Phylogenetic analysis of the sequences demonstrated that phylum Proteobacteria (52%), followed by Planctomycetes (12.7%), Chloroflexi and an unidentified bacterial group (8.8% each) were represented in the library. Deltaproteobacteria was the dominant class (62.5%) in phylum Proteobacteria with clones falling in orders Syntrophobacterales and Desulfovibrionales. Few minor phylogenetic groups, corresponding to Spirochetes, Firmicutes, Acidobacteria and Verrucomicrobia were found. Unidentified candidate groups falling in OP11, OP8 and OP3 were represented by 0.9, 2.9 and 3.9%, respectively and two clusters of the cloned sequences in this study showed very low identity to known sequences. This is the first report that discusses the phylogenetic groups in the OMZ sediments of eastern Arabian Sea individually and compares it with available data from marine hypoxic locales and water mass. LIBSHUFF statistics revealed high richness of the bacterial community of the Arabian Sea OMZ (AS-OMZ) compared to the other regions.     

Occurrence, diversity and pathogenicity of mesophilic Aeromonas in estuarine waters of the Italian coast of the Adriatic Sea

A total of 208 strains of Aeromonas were isolated by monthly sampling from two estuaries (one provided with, and the other devoid of a waste-water treatment system) on the Italian coast of the Adriatic sea between September 1994 and August 1995. Biotyping at the species level allowed the identification of 96 strains (46%) as Aer. caviae , 46 (22%) as Aer. sobria , 33 (16%) as Aer. hydrophila and 25 (12%) as Aer. veronii . Eight strains (4%) were regarded as unnamed aeromonads. Aeromonas caviae was the most prevalent species in water with a high degree of pollution, while Aer. hydrophila strains were more commonly isolated from cleaner water. Aeromonas sobria and Aer. veronii were equally distributed in both estuaries. There was no correlation between temperature and numbers of aeromonads in either estuary. Using a biochemical fingerprinting method, strains were divided into similarity groups (PhP-types) based on their biochemical phenotypes. Several different PhP-types were found in each estuary, yielding a high diversity for these strains. However, some identical PhP-types were also found in both estuaries and at different times of the year, indicating that certain Aeromonas strains can survive more widely varying physico-chemical conditions. The production of toxins capable of causing cytoskeletal-dependent changes in the morphology of Chinese hamster ovary (CHO) cells was detected in 14 strains and appeared to be dependent on the season.