Evolution of Protein Molecules

Background Improvement in lung function after macrolide antibiotic therapy has been attributed to reduction in bronchial infection by specific bacteria. However, the airway might be populated by a more diverse microbiota, and clinical features of asthma might be associated with characteristics of the airway microbiota present. Objective We sought to determine whether relationships exist between the composition of the airway bacterial microbiota and clinical features of asthma using culture-independent tools capable of detecting the presence and relative abundance of most known bacteria. Methods In this pilot study bronchial epithelial brushings were collected from 65 adults with suboptimally controlled asthma participating in a multicenter study of the effects of clarithromycin on asthma control and 10 healthy control subjects. A combination of high-density 16S ribosomal RNA microarray and parallel clone library-sequencing analysis was used to profile the microbiota and examine relationships with clinical measurements. Results Compared with control subjects, 16S ribosomal RNA amplicon concentrations (a proxy for bacterial burden) and bacterial diversity were significantly higher among asthmatic patients. In multivariate analyses airway microbiota composition and diversity were significantly correlated with bronchial hyperresponsiveness. Specifically, the relative abundance of particular phylotypes, including members of the Comamonadaceae, Sphingomonadaceae, Oxalobacteraceae, and other bacterial families were highly correlated with the degree of bronchial hyperresponsiveness. Conclusion: The composition of bronchial airway microbiota is associated with the degree of bronchial hyperresponsiveness among patients with suboptimally controlled asthma. These findings support the need for further functional studies to examine the potential contribution of members of the airway microbiota in asthma pathogenesis.

/pdf/airway-microbiota-and-bronchial-hyperresponsiveness-in-1avj6i7962.pdf

Airway microbiota and bronchial hyperresponsiveness in patients with suboptimally controlled asthma.

https://www.gastrojournal.org/article/S0016-5085(11)00922-X/pdf

Gastrointestinal Microbiome Signatures of Pediatric Patients With Irritable Bowel Syndrome

The majority of life on Earth--notably, microbial life--occurs in places that do not receive sunlight, with the habitats of the oceans being the largest of these reservoirs. Sunlight penetrates only a few tens to hundreds of meters into the ocean, resulting in large-scale microbial ecosystems that function in the dark. Our knowledge of microbial processes in the dark ocean-the aphotic pelagic ocean, sediments, oceanic crust, hydrothermal vents, etc.-has increased substantially in recent decades. Studies that try to decipher the activity of microorganisms in the dark ocean, where we cannot easily observe them, are yielding paradigm-shifting discoveries that are fundamentally changing our understanding of the role of the dark ocean in the global Earth system and its biogeochemical cycles. New generations of researchers and experimental tools have emerged, in the last decade in particular, owing to dedicated research programs to explore the dark ocean biosphere. This review focuses on our current understanding of microbiology in the dark ocean, outlining salient features of various habitats and discussing known and still unexplored types of microbial metabolism and their consequences in global biogeochemical cycling. We also focus on patterns of microbial diversity in the dark ocean and on processes and communities that are characteristic of the different habitats.

Microbial Ecology of the Dark Ocean above, at, and below the Seafloor

A dramatic exception to the general pattern of single-copy genes in bacterial and archaeal genomes is the presence of 1–15 copies of each ribosomal RNA encoding gene. The original version of the Ribosomal RNA Database (rrnDB) cataloged estimates of the number of 16S rRNA-encoding genes; the database now includes the number of genes encoding each of the rRNAs (5S, 16S and 23S), an internally transcribed spacer region, and the number of tRNA genes. The rrnDB has been used largely by microbiologists to predict the relative rate at which microbial populations respond to favorable growth conditions, and to interpret 16S rRNA-based surveys of microbial communities. To expand the functionality of the rrnDB (http://ribo some.mmg.msu.edu/rrndb/index.php), the search engine has been redesigned to allow database searches based on 16S rRNA gene copy number, specific organisms or taxonomic subsets of organisms. The revamped database also computes average gene copy numbers for any collection of entries selected. Curation tools now permit rapid updates, resulting in an expansion of the database to include data for 785 bacterial and 69 archaeal strains. The rrnDB continues to serve as the authoritative, curated source that documents the phylogenetic distribution of rRNA and tRNA genes in microbial genomes.

rrnDB: documenting the number of rRNA and tRNA genes in bacteria and archaea

A SYBR Green real-time quantitative PCR (Q-PCR) assay for the detection and quantification of Bacteria and Archaea present in the glassy rind of seafloor basalts of different ages and water depths is presented. Two sets of domain-specific primers were designed and validated for specific detection and quantification of bacterial and archaeal 16S rRNA genes in DNA extracted from basaltic glass. Total cell numbers were also estimated by fluorescence microscopy analysis of SYBR Gold-stained samples. The results from the two different approaches were concurrent, and Q-PCR results showed that the total number of cells present in basalts was in the range from 6 × 105 to 4 × 106 cells g−1 basaltic glass. Further, it was demonstrated that these cells were almost exclusively from the domain Bacteria. When applying the same methods on samples of different ages (22 years–0.1 Ma) and water depths (139–3390 mbsl), no significant differences in cell concentrations or in the relative abundance of Archaea and Bacteria were detected.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1574-6968.2008.01119.x

Enumeration of Archaea and Bacteria in seafloor basalt using real-time quantitative PCR and fluorescence microscopy

Four yellow-pigmented, Gram-negative, motile strains were isolated from the glassy rind of submarine basaltic lava from the Jan Mayen area of the Norwegian/Greenland Sea at a depth of 1300 m below sea level. The four strains had identical 16S rRNA gene sequences and were indistinguishable in all phenotypic and chemotypic tests performed, indicating that they belonged to the same species. The strains had an obligately aerobic chemo-organotrophic metabolism. The strains were capable of growth at temperatures between -2 and 34 degrees C, at pH between 6.5 and 8.6, and at sea salt concentrations between 3 and 60 g l(-1). The strains were able to utilize organic acids, amino acids and sugars but not alcohols; they were also capable of hydrolysing a wide range of macromolecules. The predominant fatty acids were 15 : 0 iso, 15 : 1 iso, 15 : 0 iso 3-OH and 17 : 0 iso 3-OH. The mean DNA G+C content of the strains was 31.4 mol%. 16S rRNA gene sequence analysis indicated that the strains were affiliated to the genera Gaetbulibacter and Algibacter. However, phenotypic characteristics, especially aerobic metabolism, suggested that the strains should be placed within a new genus. On the basis of the polyphasic characterization of the four strains, it is suggested that the strains be included in the family Flavobacteriaceae as representatives of a novel species in a new genus, for which the name Flaviramulus basaltis gen. nov., sp. nov. is proposed. The type strain is H35(T) (=CIP 109091(T)=DSM 18180(T)).

Flaviramulus basaltis gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from seafloor basalt

. Microorganisms have been reported to be associated with the alteration of the glassy margin of seafloor pillow basalts (Thorseth et al., 2001, 2003; Lysnes et al., 2004). The amount of iron and other biological important elements present in basalts and the vast abundance of basaltic glass in the earth's crust, make glass alteration an important process in global element cycling. To gain further insight into microbial communities associated with glass alteration, five microcosm experiments mimicking seafloor conditions were inoculated with seafloor basalt and incubated for one year. Mineral precipitations, microbial attachment to the glass and glass alteration were visualized by scanning electron microscopy (SEM), and the bacterial community composition was fingerprinted by PCR and denaturing gradient gel electrophoresis (DGGE) in combination with sequencing. SEM analysis revealed a microbial community with low morphological diversity of mainly biofilm associated and prosthecate microorganisms. Approximately 30 nm thick alteration rims developed on the glass in all microcosms after one year of incubation; this however was also seen in non inoculated controls. Calcium carbonate precipitates showed parallel, columnar and filamentous crystallization habits in the microcosms as well as in the sterile controls. DGGE analysis showed an alteration in bacterial community profiles in the five different microcosms, as a response to the different energy and redox regimes and time. In all microcosms a reduction in number of DGGE bands, in combination with an increase in cell abundance were recorded during the experiment. Sequence analysis showed that the microcosms were dominated by four groups of organisms with phylogenetic affiliation to four taxa: The Rhodospirillaceae, a family containing phototrophic marine organisms, in which some members are capable of heterotrophic growth in darkness and N2 fixation; the family Hyphomicrobiaceae, a group of prosthecate oligotrophic organisms; the genus Rhizobium, N2 fixating heterotrophs; and the genus Sphingomonas, which are known as bio-film producing oligotrophs. Although no bioalteration of glass could be confirmed from our experiments, oligotrophic surface adhering bacteria such as the Sphingomonas sp. and Hyphomicrobium sp. may nevertheless be important for bioalteration in nature, due to their firm attachment to glass surfaces, and their potential for biofilm production.

/pdf/microbial-colonization-and-alteration-of-basaltic-glass-182d1mdsq8.pdf

Microbial colonization and alteration of basaltic glass

Quantitative characterization of the mode and rate of phenotypic evolution is rarely applied to prokaryotes. Here, we present an analysis of temperature optimum (Topt) evolution in the thermophilic family Thermotogaceae, which has a large number of cultured representatives. We use log-rate-interval analysis to show that Topt evolution in Thermotogaceae is consistent with a Brownian motion (BM) evolutionary model. The properties of the BM model are used to a establish confidence intervals on the unknown phenotypic trait value of an uncultured organism, given its distance to a close relative with known trait value. Cross-validation by bootstrapping indicates that the predictions are robust.

Electronic supplementary material
The online version of this article (doi:10.1007/s00792-011-0381-4) contains supplementary material, which is available to authorized users.

/pdf/evolution-of-temperature-optimum-in-thermotogaceae-and-the-rr2561ppni.pdf

Evolution of temperature optimum in Thermotogaceae and the prediction of trait values of uncultured organisms

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Enumeration of Archaea and Bacteria in seafloor basalt using real-time quantitative PCR and fluorescence microscopy

Flaviramulus basaltis gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from seafloor basalt

Microbial colonization and alteration of basaltic glass

Evolution of temperature optimum in Thermotogaceae and the prediction of trait values of uncultured organisms