24 results on '"Neulinger, Sven"'
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2. Osmotic Adaptation and Compatible Solute Biosynthesis of Phototrophic Bacteria as Revealed from Genome Analyses.
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Imhoff JF, Rahn T, Künzel S, Keller A, and Neulinger SC
- Abstract
Osmotic adaptation and accumulation of compatible solutes is a key process for life at high osmotic pressure and elevated salt concentrations. Most important solutes that can protect cell structures and metabolic processes at high salt concentrations are glycine betaine and ectoine. The genome analysis of more than 130 phototrophic bacteria shows that biosynthesis of glycine betaine is common among marine and halophilic phototrophic Proteobacteria and their chemotrophic relatives, as well as in representatives of Pirellulaceae and Actinobacteria , but are also found in halophilic Cyanobacteria and Chloroherpeton thalassium . This ability correlates well with the successful toleration of extreme salt concentrations. Freshwater bacteria in general lack the possibilities to synthesize and often also to take up these compounds. The biosynthesis of ectoine is found in the phylogenetic lines of phototrophic Alpha - and Gammaproteobacteria , most prominent in the Halorhodospira species and a number of Rhodobacteraceae . It is also common among Streptomycetes and Bacilli . The phylogeny of glycine-sarcosine methyltransferase (GMT) and diaminobutyrate-pyruvate aminotransferase (EctB) sequences correlate well with otherwise established phylogenetic groups. Most significantly, GMT sequences of cyanobacteria form two major phylogenetic branches and the branch of Halorhodospira species is distinct from all other Ectothiorhodospiraceae . A variety of transport systems for osmolytes are present in the studied bacteria., Competing Interests: The authors declare no conflict of interest.
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- 2020
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3. Warming, but Not Acidification, Restructures Epibacterial Communities of the Baltic Macroalga Fucus vesiculosus With Seasonal Variability.
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Mensch B, Neulinger SC, Künzel S, Wahl M, and Schmitz RA
- Abstract
Due to ocean acidification and global warming, surface seawater of the western Baltic Sea is expected to reach an average of ∼1100 μatm p CO
2 and an increase of ∼5°C by the year 2100. In four consecutive experiments (spanning 10-11 weeks each) in all seasons within 1 year, the abiotic factors temperature (+5°C above in situ ) and p CO2 (adjusted to ∼1100 μatm) were tested for their single and combined effects on epibacterial communities of the brown macroalga Fucus vesiculosus and on bacteria present in the surrounding seawater. The experiments were set up in three biological replicates using the Kiel Outdoor Benthocosm facility (Kiel, Germany). Phylogenetic analyses of the respective microbiota were performed by bacterial 16S (V1-V2) rDNA Illumina MiSeq amplicon sequencing after 0, 4, 8, and 10/11 weeks per season. The results demonstrate ( I ) that the bacterial community composition varied in time and ( II ) that relationships between operational taxonomic units (OTUs) within an OTU association network were mainly governed by the habitat. ( III ) Neither single p CO2 nor p CO2 :Temperature interaction effects were statistically significant. However, significant impact of ocean warming was detected varying among seasons. ( IV ) An indicator OTU (iOTU) analysis identified several iOTUs that were strongly influenced by temperature in spring, summer, and winter. In the warming treatments of these three seasons, we observed decreasing numbers of bacteria that are commonly associated with a healthy marine microbial community and-particularly during spring and summer-an increase in potentially pathogenic and bacteria related to intensified microfouling. This might lead to severe consequences for the F. vesiculosus holobiont finally affecting the marine ecosystem., (Copyright © 2020 Mensch, Neulinger, Künzel, Wahl and Schmitz.)- Published
- 2020
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4. Salinity and Time Can Alter Epibacterial Communities of an Invasive Seaweed.
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Saha M, Ferguson RMW, Dove S, Künzel S, Meichssner R, Neulinger SC, Petersen FO, and Weinberger F
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The establishment of epibacterial communities is fundamental to seaweed health and fitness, in modulating ecological interactions and may also facilitate adaptation to new environments. Abiotic factors like salinity can determine bacterial abundance, growth and community composition. However, influence of salinity as a driver of epibacterial community composition (until species level) has not been investigated for seaweeds and especially under long time scales. We also do not know how abiotic stressors may influence the 'core' bacterial species of seaweeds. Following an initial (immediately after field collection) sampling of epibacterial community of an invasive red seaweed Agarophyton vermicullophylum , we conducted a long term mesocosm experiment for 5 months, to examine the influence of three different salinities (low, medium and high) at two different time points (3 months after start of experiment and 5 months, i.e., at the end of experiment) on the epibacterial community richness and composition of Agarophyton . Metagenomic sequencing showed that epibacterial communities changed significantly according to salinity and time points sampled. Epibacterial richness was significantly different between low and high salinities at both time points. Epibacterial richness also varied significantly between 3 months (after start of experiment) and 5 months (end of experiment) within low, medium and high salinity level. Irrespective of salinity levels and time points sampled 727 taxa consistently appeared in all Agarophyton samples hinting at the presence of core bacterial species on the surface of the alga. Our results indicate that both salinity and time can be major driving forces in structuring epibacterial communities of seaweeds with respect to richness and β-diversity. We highlight the necessity of conducting long term experiments allowing us to detect and understand epibacterial succession over time on seaweeds., (Copyright © 2020 Saha, Ferguson, Dove, Künzel, Meichssner, Neulinger, Petersen and Weinberger.)
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- 2020
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5. Phylogeny of Anoxygenic Photosynthesis Based on Sequences of Photosynthetic Reaction Center Proteins and a Key Enzyme in Bacteriochlorophyll Biosynthesis, the Chlorophyllide Reductase.
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Imhoff JF, Rahn T, Künzel S, and Neulinger SC
- Abstract
Photosynthesis is a key process for the establishment and maintenance of life on earth, and it is manifested in several major lineages of the prokaryote tree of life. The evolution of photosynthesis in anoxygenic photosynthetic bacteria is of major interest as these have the most ancient roots of photosynthetic systems. The phylogenetic relations between anoxygenic phototrophic bacteria were compared on the basis of sequences of key proteins of the type-II photosynthetic reaction center, including PufLM and PufH (PuhA), and a key enzyme of bacteriochlorophyll biosynthesis, the light-independent chlorophyllide reductase BchXYZ. The latter was common to all anoxygenic phototrophic bacteria, including those with a type-I and those with a type-II photosynthetic reaction center. The phylogenetic considerations included cultured phototrophic bacteria from several phyla, including Proteobacteria (138 species), Chloroflexi (five species), Chlorobi (six species), as well as Heliobacterium modesticaldum (Firmicutes) , Chloracidobacterium acidophilum (Acidobacteria), and Gemmatimonas phototrophica (Gemmatimonadetes) . Whenever available, type strains were studied. Phylogenetic relationships based on a photosynthesis tree (PS tree, including sequences of PufHLM-BchXYZ) were compared with those of 16S rRNA gene sequences (RNS tree). Despite some significant differences, large parts were congruent between the 16S rRNA phylogeny and photosynthesis proteins. The phylogenetic relations demonstrated that bacteriochlorophyll biosynthesis had evolved in ancestors of phototrophic green bacteria much earlier as compared to phototrophic purple bacteria and that multiple events independently formed different lineages of aerobic phototrophic purple bacteria, many of which have very ancient roots. The Rhodobacterales clearly represented the youngest group, which was separated from other Proteobacteria by a large evolutionary gap.
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- 2019
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6. Immediate Effects of Ammonia Shock on Transcription and Composition of a Biogas Reactor Microbiome.
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Fischer MA, Ulbricht A, Neulinger SC, Refai S, Waßmann K, Künzel S, and Schmitz RA
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The biotechnological process of biogas production from organic material is carried out by a diverse microbial community under anaerobic conditions. However, the complex and sensitive microbial network present in anaerobic degradation of organic material can be disturbed by increased ammonia concentration introduced into the system by protein-rich substrates and imbalanced feeding. Here, we report on a simulated increase of ammonia concentration in a fed batch lab-scale biogas reactor experiment. Two treatment conditions were used simulating total ammonia nitrogen concentrations of 4.9 and 8.0 g/L with four replicate reactors. Each reactor was monitored concerning methane generation and microbial composition using 16S rRNA gene amplicon sequencing, while the transcriptional activity of the overall process was investigated by metatranscriptomic analysis. This allowed investigating the response of the microbial community in terms of species composition and transcriptional activity to a rapid upshift to high ammonia conditions. Clostridia and Methanomicrobiales dominated the microbial community throughout the entire experiment under both experimental conditions, while Methanosarcinales were only present in minor abundance. Transcription analysis demonstrated clostridial dominance with respect to genes encoding for enzymes of the hydrolysis step (cellulase, EC 3.2.1.4) as well as dominance of key genes for enzymes of the methanogenic pathway (methyl-CoM reductase, EC 2.8.4.1; heterodisulfide reductase, EC 1.8.98.1). Upon ammonia shock, the selected marker genes showed significant changes in transcriptional activity. Cellulose hydrolysis as well as methanogenesis were significantly reduced at high ammonia concentrations as indicated by reduced transcription levels of the corresponding genes. Based on these experiments we concluded that, apart from the methanogenic archaea, hydrolytic cellulose-degrading microorganisms are negatively affected by high ammonia concentrations. Further, Acholeplasma and Erysipelotrichia showed lower abundance under increased ammonia concentrations and thus might serve as indicator species for an earlier detection in order to counteract against ammonia crises.
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- 2019
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7. New insights into the metabolic potential of the phototrophic purple bacterium Rhodopila globiformis DSM 161 T from its draft genome sequence and evidence for a vanadium-dependent nitrogenase.
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Imhoff JF, Rahn T, Künzel S, and Neulinger SC
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- Acetobacteraceae isolation & purification, Acetobacteraceae radiation effects, Bacterial Proteins genetics, Base Composition, Base Sequence, Chromosome Mapping, Hot Springs microbiology, Nitrogenase genetics, Photosynthetic Reaction Center Complex Proteins genetics, Photosynthetic Reaction Center Complex Proteins metabolism, Phototrophic Processes, Vanadium metabolism, Acetobacteraceae genetics, Acetobacteraceae metabolism, Bacterial Proteins metabolism, Genome, Bacterial, Nitrogenase metabolism
- Abstract
Rhodopila globiformis: is the most acidophilic anaerobic anoxygenic phototrophic purple bacterium and was isolated from a warm acidic sulfur spring in Yellowstone Park. Its genome is larger than genomes of other phototrophic purple bacteria, containing 7248 Mb with a G + C content of 67.1% and 6749 protein coding and 53 RNA genes. The genome revealed some previously unknown properties such as the presence of two sets of structural genes pufLMC for the photosynthetic reaction center genes and two types of nitrogenases (Mo-Fe and V-Fe nitrogenase), capabilities of autotrophic carbon dioxide fixation and denitrification using nitrite. Rhodopila globiformis assimilates sulfate and utilizes the C1 carbon substrates CO and methanol and a number of organic compounds, in particular, sugars and aromatic compounds. It is among the few purple bacteria containing a large number of pyrroloquinoline quinone-dependent dehydrogenases. It has extended capacities to resist stress by heavy metals, demonstrates different resistance mechanisms to antibiotics, and employs several toxin/antitoxin systems.
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- 2018
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8. Photosynthesis Is Widely Distributed among Proteobacteria as Demonstrated by the Phylogeny of PufLM Reaction Center Proteins.
- Author
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Imhoff JF, Rahn T, Künzel S, and Neulinger SC
- Abstract
Two different photosystems for performing bacteriochlorophyll-mediated photosynthetic energy conversion are employed in different bacterial phyla. Those bacteria employing a photosystem II type of photosynthetic apparatus include the phototrophic purple bacteria (Proteobacteria), Gemmatimonas and Chloroflexus with their photosynthetic relatives. The proteins of the photosynthetic reaction center PufL and PufM are essential components and are common to all bacteria with a type-II photosynthetic apparatus, including the anaerobic as well as the aerobic phototrophic Proteobacteria. Therefore, PufL and PufM proteins and their genes are perfect tools to evaluate the phylogeny of the photosynthetic apparatus and to study the diversity of the bacteria employing this photosystem in nature. Almost complete pufLM gene sequences and the derived protein sequences from 152 type strains and 45 additional strains of phototrophic Proteobacteria employing photosystem II were compared. The results give interesting and comprehensive insights into the phylogeny of the photosynthetic apparatus and clearly define Chromatiales, Rhodobacterales, Sphingomonadales as major groups distinct from other Alphaproteobacteria, from Betaproteobacteria and from Caulobacterales ( Brevundimonas subvibrioides ). A special relationship exists between the PufLM sequences of those bacteria employing bacteriochlorophyll b instead of bacteriochlorophyll a . A clear phylogenetic association of aerobic phototrophic purple bacteria to anaerobic purple bacteria according to their PufLM sequences is demonstrated indicating multiple evolutionary lines from anaerobic to aerobic phototrophic purple bacteria. The impact of pufLM gene sequences for studies on the environmental diversity of phototrophic bacteria is discussed and the possibility of their identification on the species level in environmental samples is pointed out.
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- 2018
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9. Evaluation of 16S rRNA Gene Primer Pairs for Monitoring Microbial Community Structures Showed High Reproducibility within and Low Comparability between Datasets Generated with Multiple Archaeal and Bacterial Primer Pairs.
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Fischer MA, Güllert S, Neulinger SC, Streit WR, and Schmitz RA
- Abstract
The application of next-generation sequencing technology in microbial community analysis increased our knowledge and understanding of the complexity and diversity of a variety of ecosystems. In contrast to Bacteria, the archaeal domain was often not particularly addressed in the analysis of microbial communities. Consequently, established primers specifically amplifying the archaeal 16S ribosomal gene region are scarce compared to the variety of primers targeting bacterial sequences. In this study, we aimed to validate archaeal primers suitable for high throughput next generation sequencing. Three archaeal 16S primer pairs as well as two bacterial and one general microbial 16S primer pairs were comprehensively tested by in-silico evaluation and performing an experimental analysis of a complex microbial community of a biogas reactor. The results obtained clearly demonstrate that comparability of community profiles established using different primer pairs is difficult. 16S rRNA gene data derived from a shotgun metagenome of the same reactor sample added an additional perspective on the community structure. Furthermore, in-silico evaluation of primers, especially those for amplification of archaeal 16S rRNA gene regions, does not necessarily reflect the results obtained in experimental approaches. In the latter, archaeal primer pair ArchV34 showed the highest similarity to the archaeal community structure compared to observed by the metagenomic approach and thus appears to be the appropriate for analyzing archaeal communities in biogas reactors. However, a disadvantage of this primer pair was its low specificity for the archaeal domain in the experimental application leading to high amounts of bacterial sequences within the dataset. Overall our results indicate a rather limited comparability between community structures investigated and determined using different primer pairs as well as between metagenome and 16S rRNA gene amplicon based community structure analysis. This finding, previously shown for Bacteria, was as well observed for the archaeal domain.
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- 2016
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10. Restructuring of Epibacterial Communities on Fucus vesiculosus forma mytili in Response to Elevated pCO2 and Increased Temperature Levels.
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Mensch B, Neulinger SC, Graiff A, Pansch A, Künzel S, Fischer MA, and Schmitz RA
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Marine multicellular organisms in composition with their associated microbiota-representing metaorganisms-are confronted with constantly changing environmental conditions. In 2110, the seawater temperature is predicted to be increased by ~5°C, and the atmospheric carbon dioxide partial pressure (pCO2) is expected to reach approximately 1000 ppm. In order to assess the response of marine metaorganisms to global changes, e.g., by effects on host-microbe interactions, we evaluated the response of epibacterial communities associated with Fucus vesiculosus forma mytili (F. mytili) to future climate conditions. During an 11-week lasting mesocosm experiment on the island of Sylt (Germany) in spring 2014, North Sea F. mytili individuals were exposed to elevated pCO2 (1000 ppm) and increased temperature levels (Δ+5°C). Both abiotic factors were tested for single and combined effects on the epibacterial community composition over time, with three replicates per treatment. The respective community structures of bacterial consortia associated to the surface of F. mytili were analyzed by Illumina MiSeq 16S rDNA amplicon sequencing after 0, 4, 8, and 11 weeks of treatment (in total 96 samples). The results demonstrated that the epibacterial community structure was strongly affected by temperature, but only weakly by elevated pCO2. No interaction effect of both factors was observed in the combined treatment. We identified several indicator operational taxonomic units (iOTUs) that were strongly influenced by the respective experimental factors. An OTU association network analysis revealed that relationships between OTUs were mainly governed by habitat. Overall, this study contributes to a better understanding of how epibacterial communities associated with F. mytili may adapt to future changes in seawater acidity and temperature, ultimately with potential consequences for host-microbe interactions.
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- 2016
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11. Composition of Bacterial Communities Associated with Aurelia aurita Changes with Compartment, Life Stage, and Population.
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Weiland-Bräuer N, Neulinger SC, Pinnow N, Künzel S, Baines JF, and Schmitz RA
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- Animals, Bacteria classification, Bacteria genetics, DNA, Bacterial genetics, In Situ Hybridization, Fluorescence, Microbiota, Molecular Sequence Data, Phylogeny, RNA, Ribosomal, 16S genetics, Bacteria isolation & purification, Scyphozoa growth & development, Scyphozoa microbiology
- Abstract
The scyphozoan Aurelia aurita is recognized as a key player in marine ecosystems and a driver of ecosystem change. It is thus intensely studied to address ecological questions, although its associations with microorganisms remain so far undescribed. In the present study, the microbiota associated with A. aurita was visualized with fluorescence in situ hybridization (FISH) analysis, and community structure was analyzed with respect to different life stages, compartments, and populations of A. aurita by 16S rRNA gene amplicon sequencing. We demonstrate that the composition of the A. aurita microbiota is generally highly distinct from the composition of communities present in ambient water. Comparison of microbial communities from different developmental stages reveals evidence for life stage-specific community patterns. Significant restructuring of the microbiota during strobilation from benthic polyp to planktonic life stages is present, arguing for a restructuring during the course of metamorphosis. Furthermore, the microbiota present in different compartments of the adult medusa (exumbrella mucus and gastric cavity) display significant differences, indicating body part-specific colonization. A novel Mycoplasma strain was identified in both compartment-specific microbiota and is most likely present inside the epithelium as indicated by FISH analysis of polyps, indicating potential endosymbiosis. Finally, comparison of polyps of different populations kept under the same controlled laboratory conditions in the same ambient water showed population-specific community patterns, most likely due the genetic background of the host. In conclusion, the presented data indicate that the associated microbiota of A. aurita may play important functional roles, e.g., during the life cycle., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)
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- 2015
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12. Dynamic changes of the luminal and mucosa-associated gut microbiota during and after antibiotic therapy with paromomycin.
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Heinsen FA, Knecht H, Neulinger SC, Schmitz RA, Knecht C, Kühbacher T, Rosenstiel PC, Schreiber S, Friedrichs AK, and Ott SJ
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- Adult, Bacteria genetics, Biopsy, DNA, Ribosomal chemistry, DNA, Ribosomal genetics, Healthy Volunteers, Humans, Molecular Sequence Data, Phylogeny, RNA, Ribosomal, 16S genetics, Young Adult, Anti-Bacterial Agents administration & dosage, Bacteria classification, Bacteria drug effects, Feces microbiology, Gastrointestinal Microbiome drug effects, Intestinal Mucosa microbiology, Paromomycin administration & dosage
- Abstract
Gut microbiota play a key role in the host's health system. Broad antibiotic therapy is known to disrupt the microbial balance affecting pathogenic as well as host-associated microbes. The aim of the present study was to investigate the influence of antibiotic paromomycin on the luminal and mucosa-associated microbiota at the DNA (abundance) and RNA (potential activity) level as well as to identify possible differences. The influence of antibiotic treatment on intestinal microbiota was investigated in 5 healthy individuals (age range: 20-22 years). All participants received the antibiotic paromomycin for 3 d. Fecal samples as well as sigmoidal biopsies were collected before and immediately after cessation of antibiotic treatment as well as after a recovery phase of 42 d. Compartment- and treatment status-specific indicator operational taxonomic units (OTUs) as well as abundance- and activity-specific patterns were identified by 16S rRNA and 16S rRNA gene amplicon libraries and high-throughput pyrosequencing. Microbial composition of lumen and mucosa were significantly different at the DNA compared to the RNA level. Antibiotic treatment resulted in changes of the microbiota, affecting the luminal and mucosal bacteria in a similar way. Several OTUs were identified as compartment- and/or treatment status-specific. Abundance and activity patterns of some indicator OTUs differed considerably. The study shows fundamental changes in composition of gut microbiota under antibiotic therapy at both the potential activity and the abundance level at different treatment status. It may help to understand the complex processes of gut microbiota changes involved in resilience mechanisms and on development of antibiotic-associated clinical diseases.
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- 2015
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13. Facets of diazotrophy in the oxygen minimum zone waters off Peru.
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Loescher CR, Großkopf T, Desai FD, Gill D, Schunck H, Croot PL, Schlosser C, Neulinger SC, Pinnow N, Lavik G, Kuypers MM, LaRoche J, and Schmitz RA
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- Archaea genetics, Archaea isolation & purification, Bacteria genetics, Bacteria isolation & purification, Cyanobacteria genetics, Cyanobacteria isolation & purification, Denitrification, Heterotrophic Processes, Nitrates analysis, Oceans and Seas, Oxidoreductases classification, Oxidoreductases genetics, Oxygen analysis, Peru, Phylogeny, Proteobacteria genetics, Proteobacteria isolation & purification, Seawater chemistry, Nitrogen Fixation genetics, Seawater microbiology
- Abstract
Nitrogen fixation, the biological reduction of dinitrogen gas (N2) to ammonium (NH4(+)), is quantitatively the most important external source of new nitrogen (N) to the open ocean. Classically, the ecological niche of oceanic N2 fixers (diazotrophs) is ascribed to tropical oligotrophic surface waters, often depleted in fixed N, with a diazotrophic community dominated by cyanobacteria. Although this applies for large areas of the ocean, biogeochemical models and phylogenetic studies suggest that the oceanic diazotrophic niche may be much broader than previously considered, resulting in major implications for the global N-budget. Here, we report on the composition, distribution and abundance of nifH, the functional gene marker for N2 fixation. Our results show the presence of eight clades of diazotrophs in the oxygen minimum zone (OMZ) off Peru. Although proteobacterial clades dominated overall, two clusters affiliated to spirochaeta and archaea were identified. N2 fixation was detected within OMZ waters and was stimulated by the addition of organic carbon sources supporting the view that non-phototrophic diazotrophs were actively fixing dinitrogen. The observed co-occurrence of key functional genes for N2 fixation, nitrification, anammox and denitrification suggests that a close spatial coupling of N-input and N-loss processes exists in the OMZ off Peru. The wide distribution of diazotrophs throughout the water column adds to the emerging view that the habitat of marine diazotrophs can be extended to low oxygen/high nitrate areas. Furthermore, our statistical analysis suggests that NO2(-) and PO4(3-) are the major factors affecting diazotrophic distribution throughout the OMZ. In view of the predicted increase in ocean deoxygenation resulting from global warming, our findings indicate that the importance of OMZs as niches for N2 fixation may increase in the future.
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- 2014
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14. Health- and disease-associated species clusters in complex natural biofilms determine the innate immune response in oral epithelial cells during biofilm maturation.
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Langfeldt D, Neulinger SC, Stiesch M, Stumpp N, Bang C, Schmitz RA, and Eberhard J
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- Bacteria classification, Bacteria genetics, Bacteria growth & development, Cell Line, Cluster Analysis, DNA, Bacterial chemistry, DNA, Bacterial genetics, DNA, Ribosomal chemistry, DNA, Ribosomal genetics, Gene Expression Profiling, Healthy Volunteers, Humans, Interleukins biosynthesis, Interleukins genetics, Molecular Sequence Data, Phylogeny, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, beta-Defensins biosynthesis, beta-Defensins genetics, Bacteria immunology, Biofilms growth & development, Epithelial Cells immunology, Immunity, Innate, Mouth Mucosa microbiology
- Abstract
The aim of the present study was to verify our hypothesis concerning the differential induction of various antimicrobial and immunomodulatory responses in oral epithelial cells by diverse bacterial species clusters. For this purpose, oral biofilms between 1 and 14 days of maturation (36 volunteers) were co-incubated with gingival epithelial cells. Subsequently, human β-defensin (hBD)-2, hBD-3, LL-37, interleukin (IL)-1β, IL-6, IL-8 and IL-10 mRNA expression profiles were quantified by quantitative reverse transcription PCR. The correlation between bacterial species and the host innate immune response was determined by relating these results to existing 16S rRNA phylogenetic analysis by amplicon sequencing (Langfeldt et al. 2014. PLoS One 9: e87449). Data were analysed by multiple factor analysis. Transcription of hBD-2 and hBD-3 was significantly associated with the abundance of species of the Prevotella cluster and the absence of species of the Streptococcus cluster. IL-1β, -6, -8 and -10 mRNA syntheses were significant correlated with Leptotrichia species [Leptotrichia 302H02 (0.448, P < 0.0001), Leptotrichia nbw822e09c1 (0.214, P = 0.008) and Leptotrichia wadei (0.218, P = 0.007)] of the Prevotella cluster. In the third dimension IL-10 and members of the Prevotella cluster were negatively correlated, whereas hBD-3 and IL-1β, IL-6 and IL-8 were positive correlated to axis 3, like members of the Proteobacteria cluster. In conclusion, distinct species of health- and disease-associated bacterial clusters induce antibacterial or immunomodulatory reactions in oral epithelial cells during early stages of bacteria-host interactions., (© 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.)
- Published
- 2014
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15. Salinity affects compositional traits of epibacterial communities on the brown macroalga Fucus vesiculosus.
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Stratil SB, Neulinger SC, Knecht H, Friedrichs AK, and Wahl M
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- Bacteria genetics, Biodiversity, Bacteria classification, Fucus microbiology, Salinity
- Abstract
Epibiotic biofilms have the potential to control major aspects of the biology and ecology of their hosts. Their composition and function may thus be essential for the health of the host. We tested the influence of salinity on the composition of epibacterial communities associated with the brown macroalga Fucus vesiculosus. Algal individuals were incubated at three salinities (5, 19, and 25) for 14 days and nonliving reference substrata (stones) were included in the experiment. Subsequently, the composition of their surface-associated bacterial communities was analyzed by 454 pyrosequencing of 16S rRNA gene sequences. Redundancy analysis revealed that the composition of epiphytic and epilithic communities significantly differed and were both affected by salinity. We found that 5% of 2494 epiphytic operational taxonomic units at 97% sequence similarity were responsible for the observed shifts. Epibacterial α-diversity was significantly lower at salinity 5 but did not differ between substrata. Our results indicate that salinity is an important factor in structuring alga-associated epibacterial communities with respect to composition and/or diversity. Whether direct or indirect mechanisms (via altered biotic interactions) may have been responsible for the observed shifts is discussed., (© 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.)
- Published
- 2014
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16. Effects of β-lactam antibiotics and fluoroquinolones on human gut microbiota in relation to Clostridium difficile associated diarrhea.
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Knecht H, Neulinger SC, Heinsen FA, Knecht C, Schilhabel A, Schmitz RA, Zimmermann A, dos Santos VM, Ferrer M, Rosenstiel PC, Schreiber S, Friedrichs AK, and Ott SJ
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- Ampicillin pharmacology, Cephalosporins pharmacology, Clostridioides difficile pathogenicity, Clostridium Infections drug therapy, Diarrhea drug therapy, Diarrhea microbiology, Fluoroquinolones therapeutic use, Gastrointestinal Tract microbiology, Humans, RNA, Ribosomal, 16S genetics, Sulbactam pharmacology, Clostridioides difficile drug effects, Fluoroquinolones pharmacology, Microbiota drug effects, beta-Lactams pharmacology
- Abstract
Clostridium difficile infections are an emerging health problem in the modern hospital environment. Severe alterations of the gut microbiome with loss of resistance to colonization against C. difficile are thought to be the major trigger, but there is no clear concept of how C. difficile infection evolves and which microbiological factors are involved. We sequenced 16S rRNA amplicons generated from DNA and RNA/cDNA of fecal samples from three groups of individuals by FLX technology: (i) healthy controls (no antibiotic therapy); (ii) individuals receiving antibiotic therapy (Ampicillin/Sulbactam, cephalosporins, and fluoroquinolones with subsequent development of C. difficile infection or (iii) individuals receiving antibiotic therapy without C. difficile infection. We compared the effects of the three different antibiotic classes on the intestinal microbiome and the effects of alterations of the gut microbiome on C. difficile infection at the DNA (total microbiota) and rRNA (potentially active) levels. A comparison of antibiotic classes showed significant differences at DNA level, but not at RNA level. Among individuals that developed or did not develop a C. difficile infection under antibiotics we found no significant differences. We identified single species that were up- or down regulated in individuals receiving antibiotics who developed the infection compared to non-infected individuals. We found no significant differences in the global composition of the transcriptionally active gut microbiome associated with C. difficile infections. We suggest that up- and down regulation of specific bacterial species may be involved in colonization resistance against C. difficile providing a potential therapeutic approach through specific manipulation of the intestinal microbiome.
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- 2014
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17. Composition of microbial oral biofilms during maturation in young healthy adults.
- Author
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Langfeldt D, Neulinger SC, Heuer W, Staufenbiel I, Künzel S, Baines JF, Eberhard J, and Schmitz RA
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- Humans, Species Specificity, Young Adult, Bacteria growth & development, Biofilms growth & development, Health, Mouth microbiology
- Abstract
In the present study we aimed to analyze the bacterial community structure of oral biofilms at different maturation stages in young healthy adults. Oral biofilms established on membrane filters were collected from 32 human subjects after 5 different maturation intervals (1, 3, 5, 9 and 14 days) and the respective phylogenetic diversity was analyzed by 16S rDNA amplicon sequencing. Our analyses revealed highly diverse entire colonization profiles, spread into 8 phyla/candidate divisions and in 15 different bacterial classes. A large inter-individual difference in the subjects' microbiota was observed, comprising 35% of the total variance, but lacking conspicuous general temporal trends in both alpha and beta diversity. We further obtained strong evidence that subjects can be categorized into three clusters based on three differently occurring and mutually exclusive species clusters.
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- 2014
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18. Gut microbiota disturbance during antibiotic therapy: a multi-omic approach.
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Pérez-Cobas AE, Gosalbes MJ, Friedrichs A, Knecht H, Artacho A, Eismann K, Otto W, Rojo D, Bargiela R, von Bergen M, Neulinger SC, Däumer C, Heinsen FA, Latorre A, Barbas C, Seifert J, dos Santos VM, Ott SJ, Ferrer M, and Moya A
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- Aged, Bacteria classification, Bacteria isolation & purification, Bacterial Typing Techniques methods, Biodiversity, DNA, Bacterial analysis, Feces microbiology, Gastrointestinal Tract metabolism, Gene Expression Profiling methods, Gene Expression Regulation drug effects, Humans, Male, Metabolome drug effects, RNA, Bacterial analysis, RNA, Ribosomal, 16S analysis, Anti-Bacterial Agents pharmacology, Bacteria drug effects, Gastrointestinal Tract microbiology, Microbiota drug effects, beta-Lactams pharmacology
- Abstract
Objective: Antibiotic (AB) usage strongly affects microbial intestinal metabolism and thereby impacts human health. Understanding this process and the underlying mechanisms remains a major research goal. Accordingly, we conducted the first comparative omic investigation of gut microbial communities in faecal samples taken at multiple time points from an individual subjected to β-lactam therapy., Methods: The total (16S rDNA) and active (16S rRNA) microbiota, metagenome, metatranscriptome (mRNAs), metametabolome (high-performance liquid chromatography coupled to electrospray ionisation and quadrupole time-of-flight mass spectrometry) and metaproteome (ultra high performing liquid chromatography coupled to an Orbitrap MS(2) instrument [UPLC-LTQ Orbitrap-MS/MS]) of a patient undergoing AB therapy for 14 days were evaluated., Results: Apparently oscillatory population dynamics were observed, with an early reduction in Gram-negative organisms (day 6) and an overall collapse in diversity and possible further colonisation by 'presumptive' naturally resistant bacteria (day 11), followed by the re-growth of Gram-positive species (day 14). During this process, the maximum imbalance in the active microbial fraction occurred later (day 14) than the greatest change in the total microbial fraction, which reached a minimum biodiversity and richness on day 11; additionally, major metabolic changes occurred at day 6. Gut bacteria respond to ABs early by activating systems to avoid the antimicrobial effects of the drugs, while 'presumptively' attenuating their overall energetic metabolic status and the capacity to transport and metabolise bile acid, cholesterol, hormones and vitamins; host-microbial interactions significantly improved after treatment cessation., Conclusions: This proof-of-concept study provides an extensive description of gut microbiota responses to follow-up β-lactam therapy. The results demonstrate that ABs targeting specific pathogenic infections and diseases may alter gut microbial ecology and interactions with host metabolism at a much higher level than previously assumed.
- Published
- 2013
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19. Temperature-driven shifts in the epibiotic bacterial community composition of the brown macroalga Fucus vesiculosus.
- Author
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Stratil SB, Neulinger SC, Knecht H, Friedrichs AK, and Wahl M
- Subjects
- Bacteria classification, Bacteria genetics, Biodiversity, DNA, Bacterial genetics, Denaturing Gradient Gel Electrophoresis, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Temperature, Bacteria isolation & purification, Biofilms, Fucus microbiology, Seaweed microbiology
- Abstract
The thallus surface of the brown macroalga Fucus vesiculosus is covered by a specific biofilm community. This biofilm supposedly plays an important role in the interaction between host and environment. So far, we know little about compositional or functional shifts of this epibiotic bacterial community under changing environmental conditions. In this study, the response of the microbiota to different temperatures with respect to cell density and community composition was analyzed by nonculture-based methods (denaturing gradient gel electrophoresis and 454 pyrosequencing of the 16S rRNA gene). Redundancy analysis showed that despite high variability among host individuals temperature accounted for 20% of the variation in the bacterial community composition, whereas cell density did not differ between groups. Across all samples, 4341 bacterial operational taxonomic units (OTUs) at a 97% similarity level were identified. Eight percent of OTUs were significantly correlated with low, medium, and high temperatures. Notably, the family Rhodobacteraceae increased in relative abundance from 20% to 50% with increasing temperature. OTU diversity (evenness and richness) was higher at 15 °C than at the lower and higher temperatures. Considering their known and presumed ecological functions for the host, change in the epibacterial community may entail shifts in the performance of the host alga., (© 2013 The Authors. Published by Blackwell Publishing Ltd.)
- Published
- 2013
- Full Text
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20. Unique communities of anoxygenic phototrophic bacteria in saline lakes of Salar de Atacama (Chile): evidence for a new phylogenetic lineage of phototrophic Gammaproteobacteria from pufLM gene analyses.
- Author
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Thiel V, Tank M, Neulinger SC, Gehrmann L, Dorador C, and Imhoff JF
- Subjects
- Bacterial Proteins genetics, Chile, Chromatiaceae classification, Chromatiaceae genetics, DNA Fingerprinting, DNA, Bacterial genetics, Ectothiorhodospiraceae classification, Ectothiorhodospiraceae genetics, Gammaproteobacteria classification, Genes, Bacterial, Light-Harvesting Protein Complexes genetics, Photosynthesis, Photosynthetic Reaction Center Complex Proteins genetics, Phototrophic Processes, Polymorphism, Restriction Fragment Length, Salinity, Gammaproteobacteria genetics, Phylogeny, Water Microbiology
- Abstract
Phototrophic bacteria are important primary producers of salt lakes in the Salar de Atacama and at times form visible mass developments within and on top of the lake sediments. The communities of phototrophic bacteria from two of these lakes were characterized by molecular genetic approaches using key genes for the biosynthesis of the photosynthetic apparatus in phototrophic purple bacteria (pufLM) and in green sulfur bacteria (fmoA). Terminal restriction fragment length polymorphism of the pufLM genes indicated high variability of the community composition between the two lakes and subsamples thereof. The communities were characterized by the dominance of a novel, so far undescribed lineage of pufLM containing bacteria and the presence of representatives related to known halophilic Chromatiaceae and Ectothiorhodospiraceae. In addition, the presence of BChl b-containing anoxygenic phototrophic bacteria and of aerobic anoxygenic bacteria was indicated. Green sulfur bacteria were not detected in the environmental samples, although a bacterium related to Prosthecochloris indicum was identified in an enrichment culture. This is the first comprehensive description of phototrophic bacterial communities in a salt lake of South America made possible only due to the application of the functional pufLM genes., (© 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)
- Published
- 2010
- Full Text
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21. New phylogenetic lineages of the Spirochaetes phylum associated with Clathrina species (Porifera).
- Author
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Neulinger SC, Stöhr R, Thiel V, Schmaljohann R, and Imhoff JF
- Subjects
- Animals, Bacteria genetics, Bacteria ultrastructure, DNA, Bacterial genetics, DNA, Ribosomal genetics, Molecular Sequence Data, RNA, Ribosomal, 16S genetics, Bacteria classification, Bacteria isolation & purification, Phylogeny, Porifera microbiology
- Abstract
Though spirochetes have been repeatedly found in marine sponges and other invertebrates, little attention has been paid to the specificity of this association. This study demonstrates that different geno-and morphotypes of spirochetes can reside within the same sponge individual and develop in considerable numbers. Specimens of the calcareous sponge Clathrina clathrus collected from the Adriatic Sea off Rovinj (Croatia) were found to harbor spirochete-like bacteria, which were characterized by scanning electron microscopy (SEM), 16S rRNA gene analysis, and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH). Two novel spirochete sequence types related to the Brachyspiraceae could be retrieved. By use of specifically designed CARD-FISH probes, the C. clathrus-associated sequences could be assigned to a linear and a helical spirochete morphotype. Both were located within the sponge mesohyl and resembled the spirochete-like cells identified by SEM. In addition, from a Clathrina sp., most likely C. coriacea, that originated from Indonesian coastal waters, four different spirochete type sequences were recovered. Two of these also affiliated with the Brachyspiraceae, the other two were found associated with the Spirochaetaceae, one with the genera Borrelia and Cristispira.
- Published
- 2010
- Full Text
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22. Tissue-associated "Candidatus Mycoplasma corallicola" and filamentous bacteria on the cold-water coral Lophelia pertusa (Scleractinia).
- Author
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Neulinger SC, Gärtner A, Järnegren J, Ludvigsen M, Lochte K, and Dullo WC
- Subjects
- Animals, Bacteria classification, Bacteria genetics, In Situ Hybridization, Fluorescence methods, Norway, Anthozoa microbiology, Bacteria isolation & purification
- Abstract
The cold-water coral Lophelia pertusa (Scleractinia, Caryophylliidae) is a key species in the formation of cold-water reefs, which are among the most diverse deep-sea ecosystems. It occurs in two color varieties: white and red. Bacterial communities associated with Lophelia have been investigated in recent years, but the role of the associated bacteria remains largely obscure. This study uses catalyzed reporter deposition fluorescence in situ hybridization to detect the in situ location of specific bacterial groups on coral specimens from the Trondheimsfjord (Norway). Two tissue-associated groups were identified: (i) bacteria on the host's tentacle ectoderm, "Candidatus Mycoplasma corallicola," are flasklike, pointed cells and (ii) endoderm-associated bona fide TM7 bacteria form long filaments in the gastral cavity. These tissue-bound bacteria were found in all coral specimens from the Trondheimsfjord, indicating a closer relationship with the coral compared to bacterial assemblages present in coral mucus and gastric fluid.
- Published
- 2009
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23. Phenotype-specific bacterial communities in the cold-water coral Lophelia pertusa (Scleractinia) and their implications for the coral's nutrition, health, and distribution.
- Author
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Neulinger SC, Järnegren J, Ludvigsen M, Lochte K, and Dullo WC
- Subjects
- Animals, Bacteria genetics, Cellulose metabolism, Chitin metabolism, DNA Fingerprinting, DNA, Bacterial chemistry, DNA, Bacterial genetics, DNA, Ribosomal chemistry, DNA, Ribosomal genetics, Genes, rRNA, Molecular Sequence Data, Norway, Phylogeny, Polymorphism, Restriction Fragment Length, RNA, Bacterial genetics, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Sequence Homology, Nucleic Acid, Sulfur Compounds metabolism, Anthozoa microbiology, Anthozoa physiology, Bacteria classification, Bacteria isolation & purification, Biodiversity
- Abstract
The pseudocolonial coral Lophelia pertusa (Scleractinia, Caryophylliidae) is a eurybathic, stenothermal cosmopolitan cold-water species. It occurs in two color varieties, white and red. L. pertusa builds vast cold-water coral reefs along the continental margins, which are among the most diverse deep-sea habitats. Microbiology of L. pertusa has been in scientific focus for only a few years, but the question of whether the coral holds a host-specific bacterial community has not been finally answered. Bacteria on coral samples from the Trondheimsfjord (Norway) were characterized by the culture-independent 16S rRNA gene-based techniques terminal restriction fragment length polymorphism and sequence analysis. L. pertusa revealed a high microbial richness. Clone sequences were dominated by members of the Alpha- and Gammaproteobacteria. Other abundant taxa were Bacteroidetes, Actinobacteria, Verrucomicrobia, Firmicutes, and Planctomycetes. The bacterial community of L. pertusa not only differed conspicuously from that of the environment but also varied with both the location and color variety of its host. Therefore, the microbial colonization cannot be termed "specific" sensu stricto. However, similarities to other coral-bacterium associations suggest the existence of "cold-water coral-specific" bacterial groups sensu lato. L. pertusa-associated bacteria appear to play a significant role in the nutrition of their host by degradation of sulfur compounds, cellulose, chitin, and end products of the coral's anaerobic metabolism. Some coral-associated microbes were regarded as opportunistic pathogens. Dominance of mixotrophic members of the Rhodobacteraceae in white L. pertusa could explain the wider dispersal of this phenotype by supplementary nutrition.
- Published
- 2008
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24. Spatial distribution of sponge-associated bacteria in the Mediterranean sponge Tethya aurantium.
- Author
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Thiel V, Neulinger SC, Staufenberger T, Schmaljohann R, and Imhoff JF
- Subjects
- Animals, Bacteroidetes genetics, Bacteroidetes isolation & purification, Cyanobacteria genetics, Cyanobacteria isolation & purification, Mediterranean Region, Microscopy, Electron methods, Phylogeny, Proteobacteria genetics, Proteobacteria isolation & purification, RNA, Ribosomal, 16S genetics, Bacteroidetes growth & development, Cyanobacteria growth & development, Porifera microbiology, Proteobacteria growth & development
- Abstract
The local distribution of the bacterial community associated with the marine sponge Tethya aurantium Pallas 1766 was studied. Distinct bacterial communities were found to inhabit the endosome and cortex. Clear differences in the associated bacterial populations were demonstrated by denaturing gradient gel electrophoresis (DGGE) and analysis of 16S rRNA gene clone libraries. Specifically associated phylotypes were identified for both regions: a new phylotype of Flexibacteria was recovered only from the sponge cortex, while Synechococcus species were present mainly in the sponge endosome. Light conduction via radiate spicule bundles conceivably facilitates the unusual association of Cyanobacteria with the sponge endosome. Furthermore, a new monophyletic cluster of sponge-derived 16S rRNA gene sequences related to the Betaproteobacteria was identified using analysis of 16S rRNA gene clone libraries. Members of this cluster were specifically associated with both cortex and endosome of T. aurantium.
- Published
- 2007
- Full Text
- View/download PDF
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