- Volume 137, Issue 11, 1991
Volume 137, Issue 11, 1991
- Physiology And Growth
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Autotrophic and mixotrophic growth of Gallionella ferruginea
More LessSummary: CO2 fixation and uptake of sugars by Gallionella ferruginea were demonstrated by liquid scintillation and microautoradiographic techniques. The theoretical carbon content of a G. ferruginea cell in the exponential and stationary growth phases was calculated from size measurements of images of acridine-orange-stained cells. The carbon content of a cell in the exponential phase was 1·25 × 10−14 mol and for a cell in the stationary phase it was 5 × 10−15 mol. G. ferruginea was shown to obtain all of its cell carbon from CO2 fixation when it was cultured under aerobic gradient conditions in a mineral salt solution with iron sulphide. Uptake experiments were performed with 1·6 μM-[14C]frucose, 1·6 μM-[14C]fructose and 1·3 μM-[14C]sucrose. There was significant uptake of all three sugars. Measurements of respired 14CO2 showed that 48%, 25% and 32% of the total amount of incorporated sugar was respired for glucose, fructose and sucrose, respectively. The uptake of glucose increased when the glucose concentration in the growth medium was increased. At a glucose concentration of 10 μM or higher, the cell carbon was derived exclusively from glucose, within the errors of estimation. Mixotrophic growth with 20 μM-glucose decreased the CO2 fixation to 0·4 × 10−14 mol carbon per cell, compared to autotrophically grown cells with 1·0 × 10−14 mol carbon per cell. The addition of 20 μM-glucose gave an increase in cell number in the stationary phase from 1 × 106 to 5 × 106 cells ml−1.
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- Systematics
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Membrane fatty acids as phenotypic markers in the polyphasic taxonomy of methylotrophs within the Proteobacteria
More LessSummary: A polyphasic approach to bacterial taxonomy attempts to integrate phylogenetic relationships with phenotypic marker analysis. This study describes the application of membrane fatty acids as a phenotypic marker for methylotrophs. Detailed phospholipid, ester-linked fatty acid (PLFA) profiles are reported for 17 methylotrophic eubacterial strains. These profiles included verification of double bond positions and geometries, both critical features for this analysis. Multivariate cluster analysis was used to indicate groupings of these strains along with literature values of both methylotrophs and non-methylotrophs based on the PLFA phenotype. Like many phenotypic characteristics, PLFA profiles were influenced by environmental conditions. The instabilities displayed, however, were predictable from physiological studies including increased trans/cis and cyclopropyl/cis ratios. Cluster analysis of PLFA profiles generated by separate investigators with different culture conditions indicated reproducibility by strain and species. The PLFA phenotype relationships compare favourably with phylogenetic associations based on 16S rRNA data for methylotrophs and will continue to be a valuable phenotypic marker for Proteobacteria taxonomy.
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Characterization of halotolerant Thermus isolates from shallow marine hot springs on S. Miguel, Azores
More LessSummary: Fifteen yellow-pigmented halotolerant Thermus strains from shallow marine hot springs on a beach on S. Miguel, Azores, several isolates from terrestrial hot springs on S. Miguel and mainland Portugal, and four reference strains were subjected to a numerical taxonomy study based on 61 characters by unweighted average linkage analysis (UPGMA) using the simple matching (S SM) coefficient. This analysis produced four clusters at the 75% similarity level. Two clusters, A and B, were composed of halotolerant strains from the beach hot springs and one strain isolated inland, all of which also grew at 80°C; cluster C was composed of strains from one hot spring in mainland Portugal, while cluster D was composed of terrestrial strains from the Furnas area of S. Miguel. ‘T. thermophilus’ HB-8 and Thermus B were also halotolerant, also grew at 80 °C, and formed a large group at the 61% similarity level with the strains of clusters A and B.
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Classification of medically important clostridia using restriction endonuclease site differences of PCR-amplified 16S rDNA
More LessSummary: Restriction maps were constructed of enzymically amplified 16S rRNA genes (rDNA) isolated from eight Clostridium species. Using maximum parsimony, a dendrogram was constructed from these and published 16S rRNA sequence data. Two distinct clusters were identified: cluster I contained C. difficile, C. sordelli, and C. bifermentans, and showed 30 of 35 restriction sites in common; cluster II contained C. tetani, C. perfringens C. sporogenes and C. botulinum C and G, and showed 20 of 35 restriction sites in common. Further analysis of cluster I organisms revealed that of five Hpall fragments, two were found in equal amounts in all organisms, one was found in varying amounts in all organisms, and two were found, in varying amounts, only in C. sordelli and C. bifermentans. C. sordelli-specific and C. bifermentans-specific Hpall fragments were demonstrated by Southern hybridization of rDNA. One Hpall site with in the rDNA was present on most alleles in C. bifermentans, present on a minority of alleles in C. sordelli and absent in C. difficile. This suggested that there were two 16S rRNA alleles with different sequences present within each of the genomes of C. bifermentans and C. sordelli.
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