(9/80) Roseobacter-like bacteria in red and mediterranean sea aerobic anoxygenic photosynthetic populations.

Bacteriochlorophyll a-containing aerobic anoxygenic phototrophs (AAnP) have been proposed to account for up to 11% of the total surface water microbial community and to potentially have great ecological importance in the world's oceans. Recently, environmental and genomic data based on analysis of the pufM gene identified the existence of alpha-proteobacteria as well as possible gamma-like proteobacteria among AAnP in the Pacific Ocean. Here we report on analyses of environmental samples from the Red and Mediterranean Seas by using pufM as well as the bchX and bchL genes as molecular markers. The majority of photosynthesis genes retrieved from these seas were related to Roseobacter-like AAnP sequences. Furthermore, the sequence of a novel photosynthetic operon organization from an uncultured Roseobacter-like bacterial artificial chromosome retrieved from the Red Sea is described. The data show the presence of Roseobacter-like bacteria in Red and Mediterranean Sea AAnP populations in the seasons analyzed.  (+info)

(10/80) Nereida ignava gen. nov., sp. nov., a novel aerobic marine alpha-proteobacterium that is closely related to uncultured Prionitis (alga) gall symbionts.

A Gram-negative, slightly halophilic, non-pigmented, strictly aerobic, chemo-organotrophic bacterium was isolated from Mediterranean sea water off the Spanish coast near Valencia. This strain was poorly reactive, being unable to grow in most carbon sources analysed in minimal medium. However, good growth was observed when more complex media and longer incubation times were used. Phylogenetic analysis based on an almost complete 16S rRNA gene sequence placed strain 2SM4(T) within the Roseobacter group, in the vicinity of uncultured bacteria described as gall symbionts of several species of the red alga Prionitis. Sequence similarity values between strain 2SM4(T) and the closest neighbouring species were below 95.0 %. The cellular fatty acid composition of the Mediterranean strain confirmed its position within the 'Alphaproteobacteria', sharing 18 : 1omega7c as the major cellular fatty acid. The phylogenetic distance from any taxon with a validly published name and also a number of distinguishing features support the designation of strain 2SM4(T) as representing a novel genus and species, for which the name Nereida ignava gen. nov., sp. nov. is proposed. The type strain is 2SM4(T) (=CECT 5292(T)=DSM 16309(T)=CIP 108404(T)=CCUG 49433(T)).  (+info)

(11/80) Competitive interactions in mixed-species biofilms containing the marine bacterium Pseudoalteromonas tunicata.

Pseudoalteromonas tunicata is a biofilm-forming marine bacterium that is often found in association with the surface of eukaryotic organisms. It produces a range of extracellular inhibitory compounds, including an antibacterial protein (AlpP) thought to be beneficial for P. tunicata during competition for space and nutrients on surfaces. As part of our studies on the interactions between P. tunicata and the epiphytic bacterial community on the marine plant Ulva lactuca, we investigated the hypothesis that P. tunicata is a superior competitor compared with other bacteria isolated from the plant. A number of U. lactuca bacterial isolates were (i) identified by 16S rRNA gene sequencing, (ii) characterized for the production of or sensitivity to extracellular antibacterial proteins, and (iii) labeled with a fluorescent color tag (either the red fluorescent protein DsRed or green fluorescent protein). We then grew single- and mixed-species bacterial biofilms containing P. tunicata in glass flow cell reactors. In pure culture, all the marine isolates formed biofilms containing microcolony structures within 72 h. However, in mixed-species biofilms, P. tunicata removed the competing strain unless its competitor was relatively insensitive to AlpP (Pseudoalteromonas gracilis) or produced strong inhibitory activity against P. tunicata (Roseobacter gallaeciensis). Moreover, biofilm studies conducted with an AlpP- mutant of P. tunicata indicated that the mutant was less competitive when it was introduced into preestablished biofilms, suggesting that AlpP has a role during competitive biofilm formation. When single-species biofilms were allowed to form microcolonies before the introduction of a competitor, these microcolonies coexisted with P. tunicata for extended periods of time before they were removed. Two marine bacteria (R. gallaeciensis and P. tunicata) were superior competitors in this study. Our data suggest that this dominance can be attributed to the ability of these organisms to rapidly form microcolonies and their ability to produce extracellular antibacterial compounds.  (+info)

(12/80) Phylogenetic diversity and specificity of bacteria closely associated with Alexandrium spp. and other phytoplankton.

While several studies have suggested that bacterium-phytoplankton interactions have the potential to dramatically influence harmful algal bloom dynamics, little is known about how bacteria and phytoplankton communities interact at the species composition level. The objective of the current study was to determine whether there are specific associations between diverse phytoplankton and the bacteria that co-occur with them. We determined the phylogenetic diversity of bacterial assemblages associated with 10 Alexandrium strains and representatives of the major taxonomic groups of phytoplankton in the Gulf of Maine. For this analysis we chose xenic phytoplankton cultures that (i) represented a broad taxonomic range, (ii) represented a broad geographic range for Alexandrium spp. isolates, (iii) grew under similar cultivation conditions, (iv) had a minimal length of time since the original isolation, and (v) had been isolated from a vegetative phytoplankton cell. 16S rRNA gene fragments of most Bacteria were amplified from DNA extracted from cultures and were analyzed by denaturing gradient gel electrophoresis and sequencing. A greater number of bacterial species were shared by different Alexandrium cultures, regardless of the geographic origin, than by Alexandrium species and nontoxic phytoplankton from the Gulf of Maine. In particular, members of the Roseobacter clade showed a higher degree of association with Alexandrium than with other bacterial groups, and many sequences matched sequences reported to be associated with other toxic dinoflagellates. These results provide evidence for specificity in bacterium-phytoplankton associations.  (+info)

(13/80) Roseovarius crassostreae sp. nov., a member of the Roseobacter clade and the apparent cause of juvenile oyster disease (JOD) in cultured Eastern oysters.

An alpha-proteobacterium has been identified which is believed to be the causative agent of juvenile oyster disease (JOD). Since its first isolation in 1997, the bacterium has been recovered as the numerically dominant species from JOD-affected animals throughout the north-eastern United States (Maine, New York and Massachusetts). Colonies are usually beige to pinkish-beige, although the majority of isolates recovered in 2003 from an epizootic in Martha's Vineyard, Massachusetts, produce colonies with a greenish-yellow appearance. The cells are Gram-negative, aerobic, strictly marine and rod or ovoid in appearance. They are actively motile by one or two flagella, but cells are also observed to produce tufts of polar fimbriae. The principal fatty acid in whole cells is C(18:1)omega7c and other characteristic fatty acids are C(16:0), C(10:0) 3-OH, 11-methyl C(18:1)omega7c and C(18:0). Almost without exception, isolates have 16S rRNA gene sequences that are 100% identical to each other. Phylogenetic analyses place the organism within the Roseobacter clade of the alpha-Proteobacteria, with moderate bootstrap support for inclusion in the genus Roseovarius. DNA-DNA relatedness values from pairwise comparisons of this organism with the type species of the genus (Roseovarius tolerans) and the only other described species in this genus, Roseovarius nubinhibens, were 11 and 47%, respectively. Phenotypic and biochemical dissimilarities also support the assignment of this bacterium to a novel species. The name Roseovarius crassostreae sp. nov. is proposed, with the type strain CV919-312(T) (=ATCC BAA-1102(T)=DSM 16950(T)).  (+info)

(14/80) Diurnal variation of cell proliferation in three bacterial taxa from coastal North Sea waters.

Pulse-labeling with bromodeoxyuridine (BrdU) in combination with fluorescence in situ hybridization was applied to quantify the percentage of proliferating cells in coastal North Sea waters. In order to assess diurnal variability, we sampled eight or nine times, respectively, within 3 consecutive days at two seasons. Bacteria affiliated with the Roseobacter, SAR86, and NOR5 lineages constituted on average 19% +/- 3%, 8% +/- 2%, and 6% +/- 1% of all cells in May 2002 and 17% +/- 3%, 10% +/- 2%, and 11% +/- 3% in August. The relative abundances of the three populations either remained stable, or they changed very gradually during the observation periods. On average, 38 and 39% of all Bacteria exhibited DNA de novo synthesis in May and August, respectively. The fractions of proliferating cells in bacteria of the SAR86 (May, 59%; August, 72%) and the Roseobacter (48 and 53%) lineages were significantly above the community average. A substantial cell proliferation of population NOR5 (34%) was only encountered in August, concomitant with a dinoflagellate bloom. Significant short-term fluctuations of DNA-synthesizing cells were observed in Roseobacter during May and in NOR5 during August, hinting at a pronounced (temporal or spatial) mesoscale patchiness of growth rates in these populations. Since the BrdU proliferation assay is susceptible to misinterpretation, we also modeled the expected number of labeled cells at increasing BrdU incubation times in a slowly growing bacterial population. We suggest that the absence of visible DNA synthesis in marine bacterioplankton cells after DNA pulse-labeling must not be interpreted as an indication of cell "inactivity."  (+info)

(15/80) Ecology, inhibitory activity, and morphogenesis of a marine antagonistic bacterium belonging to the Roseobacter clade.

Roseobacter strain 27-4 has been isolated from a turbot larval rearing unit and is capable of reducing mortality in turbot egg yolk sac larvae. Here, we demonstrate that the supernatant of Roseobacter 27-4 is lethal to the larval pathogens Vibrio anguillarum and Vibrio splendidus in a buffer system and inhibited their growth in marine broth. Liquid chromatography (LC) with both UV spectral detection and high-resolution mass spectrometry (HR-MS) identified the known antibacterial compound thiotropocin or its closely related precursor tropodithietic acid in the bioactive fractions. Antibacterial activity correlated with the appearance of a brownish pigment and was only formed in marine broth under static growth conditions. A thick biofilm of multicellular star-shaped aggregated cells formed at the air-liquid interface under static growth conditions. Here, the bioactive compound was the base peak in the LC-UV chromatograms of the extracts where it constituted 15% of the total peak area. Aerated conditions results in 10-fold-higher cell yield, however, cultures were nonpigmented, did not produce antibacterial activity, and grew as single cells. Production of antibacterial compounds may be quorum regulated, and we identified the acylated homoserine lactone (3-hydroxy-decanoyl homoserine lactone) from cultures of Roseobacter 27-4 using LC-HR-MS. The signal molecule was primarily detected in stagnant cultures. Roseobacter 27-4 grew between 10 and 30 degrees C but died rapidly at 37 degrees C. Also, the antibacterial compounds was sensitive to heat and was inactivated at 37 degrees C in less than 2 days and at 25 degrees C in 8 days. Using Roseobacter 27-4 as a probiotic culture will require that is be established in stagnant or adhered conditions and, due to the temperature sensitivity of the active compound, constant production must be ensured.  (+info)

(16/80) Roseovarius mucosus sp. nov., a member of the Roseobacter clade with trace amounts of bacteriochlorophyll a.

Among a group of marine isolates that were found to be positive for genes of the bacterial photosynthetic reaction centre, a strain was selected for characterization that was phylogenetically close to the genus Roseovarius. The strain, designated DFL-24T, originated from a culture of Alexandrium ostenfeldii (dinoflagellate) and contained small amounts of bacteriochlorophyll a, corresponding to about 1 % of the amount found in intensely pigmented aerobic phototrophs such as Roseobacter litoralis. Cells were rods of 0.5-0.7x1.3-3.0 microm often with uneven ends, suggesting a budding mode of division. True motility was not observed. Electron micrographs of ultrathin sections revealed a Gram-negative cell-wall structure. Cultures did not grow without addition of sea salts and tolerated up to 10 % (w/v) sea-water salts. Acetate, butyrate, tricarboxylic acid cycle intermediates, glutamate and glycerol were used as growth substrates, but not glucose or fructose. Biotin, thiamine and nicotinic acid were required as growth factors. The DNA G+C content was 63 mol%. 16S rRNA gene sequence analysis placed the strain within the Roseobacter lineage of the 'Alphaproteobacteria'. Its closest phylogenetic neighbour was Roseovarius tolerans showing 96.4 % 16S rRNA gene sequence similarity. Based also on its physiological and biochemical characteristics, the strain is considered to represent a novel species of the genus Roseovarius, Roseovarius mucosus sp. nov. Strain DFL-24T (=DSM 17069T=NCIMB 14077T) is the type strain. The 16S rRNA gene sequence of DFL-24T was found to have a similarity of 99.7 % with an unidentified strain, 253-13 (=DSM 17070), which was likewise characterized and shown to constitute another strain of the species.  (+info)