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141.
Moeller R Horneck G Rettberg P Mollenkopf HJ Stackebrandt E Nicholson WL 《Current microbiology》2006,53(3):227-231
RNA was extracted from dormant and germinating Bacillus subtilis 168 spores (intact spores and chemically decoated spores) by using rapid rupture followed by acid–phenol extraction. Spore
germination progress was monitored by assaying colony forming ability before and after heat shock and by reading the optical
density at 600 nm. The purity, yield, and composition of the extracted RNA were determined spectrophotometrically from the
ratio of absorption at 260 nm to that at 280 nm; in a 2100 BioAnalyzer, giving the RNA yield/108 spores or cells and the distribution pattern of rRNA components. The method reported here for the extraction of RNA from
dormant spores, as well as during different phases of germination and outgrowth, has proven to be fast, efficient and simple
to handle. RNA of a high purity was obtained from dormant spores and during all phases of germination and growth. There was
a significant increase in RNA yield during the transition from dormant spores to germination and subsequent outgrowth. Chemically
decoated spores were retarded in germination and outgrowth compared with intact spores, and less RNA was extracted; however,
the differences were not significant. This method for RNA isolation of dormant, germinating, and outgrowing bacterial endospores
is a valuable prerequisite for gene expression studies, especially in studies on the responses of spores to hostile environmental
conditions. 相似文献
142.
143.
Genome Organization and Localization of the pufLM Genes of the Photosynthesis Reaction Center in Phylogenetically Diverse Marine Alphaproteobacteria 总被引:2,自引:0,他引:2
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Silke Pradella Martin Allgaier Christa Hoch Orsola Puker Erko Stackebrandt Irene Wagner-Dbler 《Applied microbiology》2004,70(6):3360-3369
Genome organization, plasmid content and localization of the pufLM genes of the photosynthesis reaction center were studied by pulsed-field gel electrophoresis (PFGE) in marine phototrophic Alphaproteobacteria. Both anaerobic phototrophs (Rhodobacter veldkampii and Rhodobacter sphaeroides) and strictly aerobic anoxygenic phototrophs from the Roseobacter-Sulfitobacter-Silicibacter clade (Roseivivax halodurans, Roseobacter litoralis, Staleya guttiformis, Roseovarius tolerans, and five new strains isolated from dinoflagellate cultures) were investigated. The complete genome size was estimated for R. litoralis DSM6996T to be 4,704 kb, including three linear plasmids. All strains contained extrachromosomal elements of various conformations (linear or circular) and lengths (between 4.35 and 368 kb). In strain DFL-12, a member of a putative new genus isolated from a culture of the toxic dinoflagellate Prorocentrum lima, seven linear plasmids were found, together comprising 860 kb of genetic information. Hybridization with probes against the pufLM genes of the photosynthesis gene cluster after Southern transfer of the genomic DNAs showed these genes to be located on a linear plasmid of 91 kb in R. litoralis and on a linear plasmid of 120 kb in S. guttiformis, theoretically allowing their horizontal transfer. In all other strains, the pufLM genes were detected on the bacterial chromosome. The large number and significant size of the linear plasmids found especially in isolates from dinoflagellates might account for the metabolic versatility and presumed symbiotic association with eukaryotic hosts in these bacteria. 相似文献
144.
The sensitivity, specificity, positive and negative predictive values of a reagent strip test for the diagnosis of schistosomiasis mansoni by detecting circulating cathodic antigen (CCA) in urine were evaluated using 184 stool and urine samples collected from schoolchildren living in relatively low endemic area of schistosomiasis mansoni in Ethiopia. A combined result of stool samples processed by Kato and formol-ether concentration methods was used as gold standard. The results showed that detection of CCA in urine using reagent strip test was slightly higher than the combined results of the stool techniques (65.2 % vs 42.4 %, p > 0.05) in suggesting the prevalence of the disease. The sensitivity, specificity, positive and negative predictive values of the reagent strip test were 76.9 %, 43.4 %, 50 % and 71.9 %, respectively. The result of egg counts using Kato method suggested that detection of urine CCA could be used to indicate the intensity of infection. Nevertheless, like that of stool examination, the reagent strip test was found to be less sensitive in case of light to moderate infections. About 23.1 % of the study children who were excreting the eggs of the parasite were found negative by the reagent strip test. The relative insensitivity of a reagent strip test in low intensity of infection necessitates for the development of more sensitive assay that can truly discriminate schistosome-infected from non-infected individuals. 相似文献
145.
Stackebrandt E Frühling A Cousin S Brambilla E Lünsdorf H Verbarg S 《Current microbiology》2008,56(4):298-305
A single strain, designated BF49T, was isolated from a biofilm of a tufa deposit from the Westerhöfer rivulet, Lower Saxony, Germany. The G+C content of the genomic DNA of strain BF49T was 69 mol% and the predominant ubiquinone was Q-8. Major fatty acids were C16:1ω7c/15 iso 2OH and C16:0. Comparative 16S rRNA gene sequence analysis indicated that the isolate was placed within the genus Methylibium, class Betaproteobacteria, distantly related to the type strain Methylibium petroleiphilum LMG 22953T (97.4% similarity), Methylibium fulvum Gsoil 322T (96%), and Methylibium aquaticum IMCC1728T (95.7%). On the basis of phylogenetic and phenotypic distinctness we propose a novel species, Methylibium subsaxonicum sp. nov., with strain BF49T (DSM 19570T, CIP 109700T) as the type strain. 相似文献
146.
Microbial community dynamics in Mediterranean nutrient-enriched seawater mesocosms: changes in the genetic diversity of bacterial populations 总被引:18,自引:0,他引:18
147.
Sokolova TG Jeanthon C Kostrikina NA Chernyh NA Lebedinsky AV Stackebrandt E Bonch-Osmolovskaya EA 《Extremophiles : life under extreme conditions》2004,8(4):317-323
From 24 samples of hydrothermal venting structures collected at the East Pacific Rise (13°N), 13 enrichments of coccoid cells were obtained which grew on CO, producing H2 and CO2 at 80°C. A hyperthermophilic archaeon capable of lithotrophic growth on CO coupled with equimolar production of H2 was isolated. Based on its 16S rRNA sequence analysis, this organism was affiliated with the genus Thermococcus. Other strains of Thermococcales species (Pyrococcus furiosus, Thermococcus peptonophilus, T. profundus, T. chitonophagus, T. stetteri, T. gorgonarius, T. litoralis, and T. pacificus) were shown to be unable to grow on CO. Searches in sequence databases failed to reveal deposited sequences of genes related to CO metabolism in Thermococcales. Our work provides the first evidence of anaerobic CO oxidation coupled with H2 production performed by an archaeon as well as the first documented case of lithotrophic growth of a Thermococcales representative. 相似文献
148.
Anesti V Vohra J Goonetilleka S McDonald IR Sträubler B Stackebrandt E Kelly DP Wood AP 《Environmental microbiology》2004,6(8):820-830
This is the first study to demonstrate that diverse methylotrophic bacteria occur in the human foot microflora. Polymerase chain reaction (PCR) amplification of DNA from the soles and toe clefts of feet of five subjects indicated Methylobacterium strains to be present in all cases. Polymerase chain reaction amplification also showed the gene for the alpha-subunit of methanol dehydrogenase (mxaF) to be present in all samples. Two types of mxaF were recovered, one closest to that of Methylobacterium extorquens and the other most similar to that of Hyphomicrobium methylovorum. Numerous methylotrophic strains able to grow on methylamine were isolated with ease from the feet of nine volunteers. These were found by 16S rRNA analysis to be most closely related to Methylobacterium species, Brevibacterium casei, Pseudomonas strain NZ099 and P. migulae. Three strains from two subjects were of a novel species, Methylobacterium podarium sp. nov. This facultatively methylotrophic, obligately aerobic, pink-pigmented, non-motile rod grew with a wide range of multicarbon and one-carbon compounds including citrate, xylose, mono-, di-, and trimethylamine, dimethylsulphide, methanethiol, dimethylsulphoxide, dimethylsulphone and methanol. 相似文献
149.
Tindall BJ Brambilla E Steffen M Neumann R Pukall R Kroppenstedt RM Stackebrandt E 《Environmental microbiology》2000,2(3):310-318
Rapid and inexpensive sorting of bacterial isolates may be achieved using Fourier transform infrared spectroscopy (FT-IR), a method that has hitherto been applied to identification and classification. The comprehensive characterization of environmental samples requires the isolation of large numbers of isolates using different growth media and growth conditions. In such cases, sorting the isolates is critical before isolates are subjected to more detailed studies. Using FT-IR, isolates are grown under standardized conditions, and 100 strains can be tested within less than 8 h. Chemotaxonomic and molecular characterization of members of clusters emerging from FT-IR analysis either at a level of spectral distance values below 20–30 (analysis of region 600–800 cm−1 , average linkage algorithm) or at spectral heterogeneity values below 75 (regions 1200–900, 3000–2798 and 901–698, scaling to first region, Ward's algorithm) reveals great similarities in fatty acids and 16S rDNA sequences. As judged from riboprinting analyses and fatty acid analyses, FT-IR analysis is able to unravel intraspecific subclustering. The example used in this study of 100 isolates from a mat system, Lake Fryxell, Dry Valleys, Antarctica, selected from a larger number of isolates, picked mainly on the basis of colony pigmentation and form, reveals the utility of the method for identifying the number of putative species quickly. The method described is able to select strains rapidly that represent clusters at the specific and intraspecific level for subsequent characterization. 相似文献
150.
Isolation of tellurite- and selenite-resistant bacteria from hydrothermal vents of the Juan de Fuca Ridge in the Pacific Ocean 总被引:1,自引:0,他引:1
Rathgeber C Yurkova N Stackebrandt E Beatty JT Yurkov V 《Applied and environmental microbiology》2002,68(9):4613-4622
Deep-ocean hydrothermal-vent environments are rich in heavy metals and metalloids and present excellent sites for the isolation of metal-resistant microorganisms. Both metalloid-oxide-resistant and metalloid-oxide-reducing bacteria were found. Tellurite- and selenite-reducing strains were isolated in high numbers from ocean water near hydrothermal vents, bacterial films, and sulfide-rich rocks. Growth of these isolates in media containing K(2)TeO(3) or Na(2)SeO(3) resulted in the accumulation of metallic tellurium or selenium. The MIC of K(2)TeO(3) ranged from 1,500 to greater than 2,500 micro g/ml, and the MIC of Na(2)SeO(3) ranged from 6,000 to greater than 7,000 micro g/ml for 10 strains. Phylogenetic analysis of 4 of these 10 strains revealed that they form a branch closely related to members of the genus Pseudoalteromonas, within the gamma-3 subclass of the Proteobacteria. All 10 strains were found to be salt tolerant, pH tolerant, and thermotolerant. The metalloid resistance and morphological, physiological, and phylogenetic characteristics of newly isolated strains are described. 相似文献