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Re-evaluation of the status of the genus Oerskovia, reclassification of Promicromonospora enterophila (Jáger et al. 1983) as Oerskovia enterophila comb. nov. and description of Oerskovia jenensis sp. nov. and Oerskovia paurometabola sp. nov.

Phylogenetic analysis of Promicromonospora enterophila indicates that this taxon clusters with Cellulomonas species, adjacent to Cellulomonas turbata (basonym Oerskovia turbata). 16S rDNA analysis, DNA-DNA reassociation, riboprinting, peptidoglycan analysis and determination of phenotypic properties of various strains of P. enterophila and C turbata reveal that they form a cluster that can be distinguished unambiguously from other Cellulomonas species by morphology, amino acid composition of the cell wall and 16S rDNA signatures. As a result of thispolyphasic study, it appears taxonomically reasonable to re-establish the genus Oerskovia for C turbata and to reclassify P. enterophila as Oerskovia enterophila comb. nov.; two novel species, Oerskovia jenensis sp. nov. (type strain DSM 46000T = CIP 100330T) and Oerskovia paurometabola sp. nov. (type strain DSM 14281T = LMG 20385T), are also proposed.

Actinomycetales↗

A phage-resistant mutant of Lactobacillus casei which permits phage adsorption but not genome injection.

A phage-resistant mutant of Lactobacillus casei (strain YIT 9021) was capable of adsorbing both PL-1 and J-1 phages, but did not yield phage-infected cells. The mutant and wild-type strains were identical in morphology and sugar composition of the cell walls. Attempts to induce prophages from strain YIT 9021 were unsuccessful. Electron microscopic examination of negatively stained mixtures of phage PL-1 and YIT 9021 bacteria revealed that the phages were adsorbed to the cells in a tail-first orientation. All the absorbed phages had DNA-filled heads. It was concluded that PL-1 adsorbed normally but was blocked in the injection of the phage genome into the cell.

Bacteriophages↗

Bacillus barbaricus sp. nov., isolated from an experimental wall painting.

In a project concerning bacterial colonization of experimental wall paintings, a large number of isolates have been acquired with high similarities in their whole-cell protein patterns obtained after SDS-PAGE. Of this group, four strains, designated V2-BIII-A2(T), V2-BI-A9, V2-BI-04 and V2-BII-A8, were chosen for further characterization. Banding patterns obtained after ERIC-PCR were barely distinguishable among these four strains, indicating their affiliation within a single species. The isolates also displayed nearly identical biochemical and physiological features. The chemotaxonomic characteristics, including polar lipids, quinone systems, cell-wall diamino acid composition and fatty acid profiles, were in good agreement with those of numerous previously described Bacillus species. 16S rDNA analysis of strain V2-BIII-A2(T) showed that this bacterium belongs to the genus Bacillus, with highest sequence similarities to Bacillus megaterium (94.6%), Bacillus flexus (94.4%) and the alkaliphilic Bacillus cohnii (94.2%). Based on almost identical biochemical, physiological and chemotaxonomic traits, ERIC-PCR-generated genomic fingerprints and comparative 16S rDNA sequence analysis, it is demonstrated that the four isolates represent a novel species of the genus Bacillus, for which the name Bacillus barbaricus sp. nov. is proposed. The type strain is V2-BIII-A2(T) (= CCM 4982(T) = DSM 14730(T)).

Austria↗

rpoB gene sequence-based characterization of emerging non-tuberculous mycobacteria with descriptions of Mycobacterium bolletii sp. nov., Mycobacterium phocaicum sp. nov. and Mycobacterium aubagnense sp. nov.

Over the past 10 years, 16S rRNA gene sequencing has contributed to the establishment of more than 45 novel species of non-tuberculous mycobacteria and to the description of emerging mycobacterial infections. Cumulative experience has indicated that this molecular tool underestimates the diversity of this group and does not distinguish between all recognized mycobacterial taxa. In order to improve the recognition of emerging rapidly growing mycobacteria (RGM), rpoB gene sequencing has been developed. Our previous studies have shown that an RGM isolate is a member of a novel species if it exhibits >3 % sequence divergence in the rpoB gene from the type strains of established species. When applied to a collection of 59 clinical RGM isolates, rpoB gene sequencing revealed nine novel isolates (15.3 %) whereas only two isolates (3.4 %) were deemed to be novel by conventional 16S rRNA gene sequence analysis. A polyphasic approach, including biochemical tests, antimicrobial susceptibility analyses, hsp65, sodA and recA gene sequence analysis, DNA G+C content determination and cell-wall fatty acid composition analysis, supported the evidence that these nine isolates represent three novel species. Whereas Mycobacterium phocaicum sp. nov. (type strain N4T = CIP 108542T = CCUG 50185T) and Mycobacterium aubagnense sp. nov. (type strain U8T = CIP 108543T = CCUG 50186T; Mycobacterium mucogenicum group) were susceptible to most antibiotics, Mycobacterium bolletii sp. nov. (type strain BD(T) = CIP 108541T = CCUG 50184T; Mycobacterium chelonae-abscessus group) was resistant to the quinolones, tetracycline, macrolides and imipenem. Only M. bolletii was resistant to clarithromycin. These data illustrate that rpoB gene sequence-based identification is a powerful tool to characterize emerging RGM and mycobacterial infections and provides valuable help in differentiating RGM at both the intra- and interspecies level, thus contributing to a faster and more efficient diagnosis and epidemiological follow-up.

Anti-Bacterial Agents↗

Differential Localization of Carbohydrate Epitopes in Plant Cell Walls in the Presence and Absence of Arbuscular Mycorrhizal Fungi.

Two monoclonal antibodies (McAbs) generated against rhamnogalacturonan I and characterized as specific for a terminal [alpha]-(1->2)-linked fucosyl-containing epitope (CCRC-M1) and for an arabinosylated [beta]-(1,6)-galactan epitope (CCRC-M7) were used in immunogold experiments to determine the distribution of the epitopes in four plants. Allium porrum, Zea mays, Trifolium repens, and Nicotiana tabacum plants were chosen as representatives of monocots and dicots with different wall structures. Analyses were performed on root tissues in the presence and absence of arbuscular mycorrhizal fungi. A differential localization of the two cell wall epitopes was found between tissues and between species: for example, in leek, CCRC-M1 labeled epidermal and hypodermal cells, whereas CCRC-M7 labeled cortical cells only. Clover walls were labeled by both McAbs, whereas maize and tobacco were only labeled by CCRC-M7. In the presence of the arbuscular mycorrhizal fungi, labeling was additionally found in an apoplastic compartment typical of the symbiosis (the interface) occurring around the intracellular hyphae. Epitopes binding both McAbs were found in the interfacial material, and their distribution mirrored the pattern found in the host cell wall. These findings demonstrate that the composition of the interface zone in a fungus-plant symbiosis reflects the composition of the wall of the host cell.

Journal Article↗

Requirement of ginkgo pollen-derived tissue cultures for boron and effects of boron deficiency.

Ginkgo biloba L. pollen-derived tissue, which is made up of small, friable masses of homogeneous parenchymatous cells, was shown to require boron in the culture medium. If no boron is supplied, growth soon stops. Growth responses to additions of boron were observed up to an optimum level of 0.1 mg of boron per liter.Histological examination and chemical analyses showed 2 general effects of boron deficiency: (1) a reduced rate of cell division, with no significant effect on cell size, and (2) some alteration in composition of the cell walls. With the exception of a reduction in fructose, the concentration of soluble and of readily hydrolyzable carbohydrates, and the concentration of protein in the tissue, were not affected by boron deficiency.

Journal Article↗

Serological diversity within the species Legionella spiritensis.

A strain of Legionella isolated from the environment which could not initially be identified was shown by restriction fragment length polymorphisms to be a Legionella spiritensis. This was confirmed by DNA homology studies, cell wall fatty acid composition and isoprenoid quinone analysis. This strain, which is only the second reported representative of the species, was shown to be serologically distinct from the type strain of L. spiritensis and all other serogroups of Legionella.

Antigenic Variation↗

Transcript profiling of Eucalyptus xylem genes during tension wood formation.

Tension wood formed in response to gravitational force is a striking example of the plasticity of angiosperm wood. In this study our goal was to characterize the early changes in gene expression during tension wood formation in Eucalyptus. Using cDNA array technology, transcript profiling of 231 genes preferentially expressed in differentiating Eucalyptus xylem was followed from 6 h to 1 wk of a tension time course of artificially bent Eucalyptus trees. 196 genes were differentially regulated between control and bent trees, some exhibiting distinctive expression patterns related to changes in secondary cell wall structure and composition. For instance, expression of a cellulose synthase gene was well correlated with the appearance of the G-layers. Cluster correlation analysis revealed differential regulation of lignin biosynthetic genes and may also be used to help infer the function of unknown gene products. Eucalyptus wood transcriptome analysis during tension wood formation not only provided new clues into the transcriptional regulatory network of genes preferentially expressed in xylem, but also highlighted candidate genes responsible for the genetic and environmentally induced variation of wood quality traits.

Biomechanical Phenomena↗

Molecular phylogeny of labyrinthulids and thraustochytrids based on the sequencing of 18S ribosomal RNA gene.

Labyrinthulids and thraustochytrids are unicellular heterotrophs, formerly considered as fungi, but presently are recognized as members in the stramenopiles of the kingdom Protista sensu lato. We determined the 18S ribosomal RNA gene sequences of 14 strains from different species of the six genera and analyzed the molecular phylogenetic relationships. The results conflict with the current classification based on morphology, at the genus and species levels. These organisms are separated, based on signature sequences and unique inserted sequences, into two major groups, which were named the labyrinthulid phylogenetic group and the thraustochytrid phylogenetic group. Although these groupings are in disagreement with many conventional taxonomic characters, they correlated better with the sugar composition of the cell wall. Thus, the currently used taxonomic criteria need serious reconsideration.

Animals↗

Addition of xylitol to the growth medium of Streptococcus mutans OMZ 176--effect on the synthesis of extractable glycerol-phosphate polymers.

Addition of 5% xylitol to growing cultures of Streptococcus mutans OMZ 176 reduced the amount of extractable glycerol-phosphate polymers in these cells compared to S. mutans cultures grown in medium with 5% glucose added. The glycerol-phosphate polymers were extracted from the cells by hot phenol-water-extraction, and separated by column chromatography. Lipoteichoic acid (LTA) was identified by indirect haemagglutination tests and lipid analysis. The amount of LTA extracted from the cells or present in the medium was not significantly different. It is suggested that the accumulation of intracellular xylitol-phosphate observed in a previous study causes an effect similar to glucose starvation, which is known to affect the composition of the cell wall.

Dental Plaque↗

Legionella sainthelensi: a new species of Legionella isolated from water near Mt. St. Helens.

Six strains of a new species, Legionella sainthelensi, were isolated from freshwater in areas affected by the volcanic eruptions of Mt. St. Helens in the state of Washington. Strains of L. sainthelensi are culturally and biochemically similar to other legionellae. They grow on buffered charcoal yeast agar but not on media that lack cysteine. They are gram-negative, nonsporeforming, motile rods that are positive in reactions for catalase, oxidase, gelatin liquefaction, and beta-lactamase. They are negative in reactions for urease, hydrolysis of hippurate, reduction of nitrates, fermentation of glucose, and blue-white autofluorescence. Their cell wall fatty acid composition is qualitatively similar to those of other legionellae, with 50 to 62% branched-chain fatty acids. They contain the isobranched-chain 14- and 16-carbon acids and anteisobranched-chain 15- and 17-carbon acids and relatively large amounts of straight-chain 16-carbon acid. All strains of L. sainthelensi contain approximately equal amounts of ubiquinones Q9, Q10, Q11, and Q12, a pattern similar to those of Legionella bozemanii, Legionella dumoffi, and Legionella longbeachae. Serological cross-reactions were observed between L. sainthelensi, both serogroups of L. longbeachae, and Legionella oakridgensis. Three strains of L. sainthelensi were greater than 90% related by DNA hybridization. The type strain of L. sainthelensi, Mt. St. Helens 4, was 36% related to the type strain of L. longbeachae and 3 to 14% related to the other nine described Legionella species.

Disasters↗

Isolation and characterization of yeast monomorphic mutants of Candida albicans.

A method was devised for the isolation of yeast monomorphic (LEV) mutants of Candida albicans. By this procedure, about 20 stable yeast-like mutants were isolated after mutagenesis with ethyl methane sulfonate. The growth rate of the mutants in different carbon sources, both fermentable and not, was indistinguishable from that of the parental strain, but they were unable to grow as mycelial forms after application of any of the common effective inducers, i.e., heat shock, pH alterations, proline addition, or use of GlcNAc as the carbon source. Studies performed with one selected strain demonstrated that it had severe alterations in the chemical composition of the cell wall, mainly in the levels of chitin and glucans, and in specific mannoproteins, some of them recognizable by specific polyclonal and monoclonal antibodies. It is suggested that these structural alterations hinder the construction of a normal hyphal wall.

Candida albicans↗

Relation of polysaccharide content to some biological properties of endotoxins from mutants of Salmonella typhimurium.

Kessel, R. W. I. (Rutgers, The State University, New Brunswick, N.J.), Henry H. Freedman, and Werner Braun. Relation of polysaccharide content to some biological properties of endotoxins from mutants of Salmonella typhimurium. J. Bacteriol. 92:592-596. 1966.-Endotoxins were extracted by the phenol-water procedure from a variety of Salmonella typhimurium mutants with known differences in the composition of their cell wall polysaccharides. The lethality of these preparations for mice proved to be correlated with the complexity of the polysaccharide: endotoxin from the smooth parent strain and from rough strains with several sugars attached to the heptose-phosphate backbone were of high toxicity, whereas endotoxin from a mutant possessing only glucose attached to the heptose-phosphate backbone was less toxic, and endotoxin from a mutant possessing the backbone only was least toxic. All of these mutants yielded endotoxins that were equally capable of protecting mice against subsequent challenge with Pseudomonas aeruginosa. Material obtained from a heptoseless mutant by the phenol-water method proved to be neither toxic nor protective. The apparent dissociation of biological properties that can be achieved with the aid of endotoxin preparations from certain mutants is discussed in terms of possible mechanisms.

Animals↗

Mycobacterium barrassiae sp. nov., a Mycobacterium moriokaense group species associated with chronic pneumonia.

Three identical isolates of new rapidly growing mycobacteria (RGM) were recovered from the bronchial aspirate and sputum from a 49-year-old woman presenting with lung lesions. The case met the American Thoracic Society criteria for the diagnosis of nontuberculous mycobacterial infection. The three isolates grew in 3 days at 24 to 42 degrees C. The 16S rRNA gene sequence analysis indicated that the sequences of the isolates were identical and shared 99.7% and 98.1% similarities with those of Mycobacterium moriokaense and Mycobacterium gadium, respectively. Partial 723-bp rpoB sequence analyses indicated that the sequences of the isolates shared 95.8% and 92.3% similarities with those of M. moriokaense and M. gadium, respectively. Polyphasic identification (including biochemical tests; antimicrobial susceptibility profiling; and hsp65, recA, and sodA gene sequence analyses, as well as G+C content determination and cell wall fatty acid composition analysis) supported the evidence that these isolates were representative of a new species. Phylogenetic analyses confirmed the close relationships of the isolates with M. moriokaense and the defined M. moriokaense group. These isolates were susceptible to the antimicrobials currently recommended for the treatment of RGM infections. These isolates differed from M. moriokaense by their susceptibility to vancomycin. We propose the name Mycobacterium barrassiae sp. nov. for this new species. The type strain is N7T (CIP 108545T and CCUG 50398T).

Anti-Bacterial Agents↗

Description of Mycobacterium conceptionense sp. nov., a Mycobacterium fortuitum group organism isolated from a posttraumatic osteitis inflammation.

A nonpigmented rapidly growing mycobacterium was isolated from wound liquid outflow, bone tissue biopsy, and excised skin tissue from a 31-year-old woman who suffered an accidental open right tibia fracture and prolonged stay in a river. The three isolates grew in 3 days at 24 to 37 degrees C. 16S rRNA sequence analyses over 1,483 bp showed that they were identical and shared 99.7% (4-bp difference) sequence similarity with that of Mycobacterium porcinum, the most closely related species. Partial rpoB (723 bp) sequence analyses showed that the isolates shared 97.0% sequence similarity with that of M. porcinum. Further polyphasic approaches, including biochemical tests, antimicrobial susceptibility analyses, and hsp65, sodA, and recA gene sequence analysis, as well as % G+C determination and cell wall fatty acid composition analysis supported the evidence that these isolates were representative of a new species. Phylogenetic analyses showed the close relationship with M. porcinum in the Mycobacterium fortuitum group. The isolates were susceptible to most antibiotics and exhibited evidence for penicillinase activity, in contrast to M. porcinum. We propose the name Mycobacterium conceptionense sp. nov. for this new species associated with posttraumatic osteitis. The type strain is D16(T) (equivalent to CIP 108544(T) and CCUG 50187(T)).

Adult↗

Controlled cell disruption: a comparison of the forces required to disrupt different micro-organisms.

A cell disrupter has been developed which can measure the forces required to disrupt both eukaryotic and prokaryotic cells. It operates a continuous process and will disrupt both large and small volumes. Shear forces are set up when a suspension under laminar flow conditions is released under high pressure through a short orifice. If the applied pressure is altered, the shear forces are simultaneously changed so that the amount of cell disruption can be compared under different known and repeatable conditions. The disrupter is now manufactured and supplied by Stansted Fluid Power Limited, Stansted, England. Phase-contrast microscopy has shown that the disrupter will break a variety of organisms including Chlorella, Aspergillus fumigatis, Fusarium sp., Saccharomyces cerevisiae, Escherichia coli, Lactobacillus casei, Bacillus subtilis, Clostridium perfringens, Streptococcus faecalis, Streptococcus zooepidermicus and Staphylococcus aureus. The cells are not all broken at one pressure but a certain pressure must be applied before disruption starts which will then increase rapidly as the applied pressure is increased. The applied pressure required to disrupt half the population in a culture is different from one species to another, rods being disrupted more easily than spheres. The case of disruption seems to be related to the shape and chemical composition of the cell wall. Furthermore, the disrupting process, in an unsynchronized culture is not random and may be related to the statistical size distribution of the cells.

Bacteria↗

Development of fern sporangia: a fluorescence microscopy study.

The utility of fluorescence microscopy for studying development of fern spores is investigated. Changes in the fluorescence characteristics during the developmental stages of fern sporangia can be attributed to the changes in the chemical composition of the cell wall. Bright blue autofluorescence of the spores indicated the presence of sporopollenin. The sporangial walls and the spores autofluoresced yellow under blue light excitation. Fluorescence microscopy is a useful addition to light, scanning, and transmission electron microscopy because living specimens can be studied owing to their fluorescence properties.

Biopolymers↗

[Review of the systematic position of Actinomyces orientalis (Pittenger et Brigham) Krass. 1970 and its reclassification as Proactinomyces orientalis (Pittenger et Brigham) comb. nov].

The micromorphology, growth cycle, chemical composition of the cell walls, and the susceptibility to lysozyme and specific phages were studied with the culture of Actinomyces orientalis RIA-1074. It is seggested on the basis of these studied to transfer the culture from the Actinomyces genus to the Proactinomyces genus and to rename it Proactinomyces orientalis (Pittinger et Brigham) comb. nov.

Actinomyces↗