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PubMed1992

Phage typing--a useful tool in actinomycete systematics.

A total of 905 strains of several genera of the Actinomycetales were tested in respect to their sensitivity to genus- and species-specific actinophages. The results provided clear evidence that phage typing is a useful aid for the identification of actinomycetes. At the genus level, the specificity of actinophages has led to the correct placement of falsely identified strains, as well as to the rearrangement of genera. At the species level, however, phage typing has been less successful: only some actinophages are available so far, lysing strains (often not all) of one or a few related species (synonyms), e.g., of the genera Streptomyces and Amycolatopsis, as well as Saccharopolyspora.

Actinomycetales↗

PubMed2004

Identification of a promoter motif regulating the major DNA damage response mechanism of Mycobacterium tuberculosis.

The principal response of many bacteria to DNA damage is mediated by a mechanism dependent on the LexA and RecA proteins. However, Mycobacterium tuberculosis was recently reported to regulate a majority of DNA repair genes independently of RecA and LexA, suggesting that an unknown RecA/LexA-independent mechanism controls the major DNA damage response pathway in this organism. Here we have identified a motif tTGTCRgtg-8nt-TAnnnT that defines a novel RecA/LexA-independent promoter (RecA-NDp) of M. tuberculosis. Furthermore, we show that the RecA-NDp type of promoter precedes DNA repair genes in other Actinomycetales.

Actinomycetales↗

PubMed2005

Biochemical evidence for the proteolytic degradation of infectious prion protein PrPsc in hamster brain homogenates by foodborne bacteria.

PrP(Sc) is a general term to describe the infectious agent causing transmissible spongiform encephalopathy (TSE), and the protease-resistant form of cellular PrP(C). In this study, we have identified several protease-secreting bacteria able to degrade PrP(Sc) under more or less native conditions (30 degrees C, pH 8), focusing on strains isolated mainly from cheese. One hundred and ninty-nine protease-secreting isolates belonging to the Actinomycetales and Bacillales were screened for the expression of PrP(Sc) degrading activity by a Western blot procedure. Only 6 strains belonging to the following species were found to exhibit such an activity: Arthrobacter nicotianae, Bacillus licheniformis, Brachybacterium conglomeratum, Brachybacterium tyrofermentans and Staphylococcus sciuri and Serratia spp. As revealed by a general protease assay based on dye-labeled Azocoll substrate, the PrP(Sc) degrading activity was not directly correlated to the total level of secreted proteolytic activity of these organisms. This indicates that specific proteases are required for the degradation of PrP(Sc). Our study also suggests the potential use of such starter bacteria or their proteases for application in PrP(Sc) degradation and decontamination under native conditions.

Actinomycetales↗

PubMed1990

Disseminated skeletal lesions associated with pathogenic actinomycetes in a Pygmy hog (Sus salvanius).

The clinical and pathological features of an unusual crippling bone disease in an adult male Pygmy Hog, Sus salvanius, born the smallest in a litter of five and representing the rarest of the known living Suidae, are described. Radiological studies revealed severe spondylosis deformans and focal sharply demarcated radiolucencies in virtually the whole skeleton, but particularly in the bones of the skull, the processes of multiple vertebrae, the ribs, scapulae and parts of the humeri and femora. The clinical chemistry measurements were indicative of pathological lytic processes in the skeleton. The focal bone lesions consisted of caseous necrosis, dystrophic calcification and peripheral fibroblastic demarcation. They contained colonies of filamentous bacteria identified as members of the Order Actinomycetales. Case history evidence suggests that the infection may have resulted from repeated skin trauma inflicted by litter-mates.

Actinomycetales Infections↗

PubMed2000

Mycolic acids from Rhodococcus, Gordonia, and Dietzia.

The mycolic acids from 11 species of Rhodococcus, seven species of Gordonia, and one species of Dietzia were analyzed using capillary gas chromatography and mass spectrometry (GLC/MS). All strains tested in this study were divided into three groups according to the degree of double bonds and the average carbon number (Av.Nc.) of their mycolic acids. The genus Gordonia belongs to the first group possessing an Av.Nc. in the upper 50s and 60s with 0 to 5 double bonds. Some Rhodococcus species possessed Av.Nc. in the 40s with a variety of distributions of polyunsaturated fatty acids from 0 to 4. The rest of the Rhodococcus species and the genus Dietzia possessed Av.Nc. in the 30s with saturated fatty acids. We previously reported on Nocardia strains whose Av.Nc. were in the 50s. Considering the identification of mycolic acid-containing Actinomycetales at the generic level, the Av.Nc. proved to be useful as a means of differentiating the genera Rhodococcus, Gordonia and Nocardia. The genus Dietzia was found to have its own characteristic constitution of mycolic acid molecular species. The mycolic acids from D. maris 58001T were characterized by an almost equal amount of constituents of even- and odd-numbered carbon chains, whereas the major components of mycolic acids in all other strains had even-numbered carbon chains. Another characteristic of Dietzia was some even-numbered mycolic acids which contained odd-numbered straight chains with odd-numbered alpha-branches. These characteristics indicated that Dietzia might possess a novel fatty acid biosynthesis system.

Actinomycetales↗

PubMed2002

Parkinson's: another look.

Recent studies in the Parkinson's literature have cited a tuberculosis-like germ called Nocardia as being responsible for Parkinson's disease. Kohbata seemingly cemented a relationship between Nocardia and Parkinson's by finding serologic evidence in 20 of 20 Parkinson's patients, acknowledging that blood tests for Nocardia and the mycobacteria such as tuberculosis often cross-react, as they belong to the same order of bacteria, the Actinomycetales. Besides this difficulty in differentiation, a well-used medical school textbook of microbiology, Atlas, points out that even among experts, different observers may classify the same strain of bacteria as Nocardia or Mycobacterium tuberculosis. Parkinson's: another look is a theoretical article which presents compelling, well-documented evidence for an infectious cause for Parkinson's disease on historical, epidemiological, pharmacologic, microbiological, and biochemical levels.

Actinomycetales Infections↗

PubMed2003

Isolation and characterisation of Nocardioides sp. SP12, an atrazine-degrading bacterial strain possessing the gene trzN from bulk- and maize rhizosphere soil.

We report the characterisation of Nocardioides sp. SP12, an atrazine-degrading bacteria isolated from atrazine-treated bulk- and maize rhizosphere soil. Based on 16S rDNA alignment, strain SP12 showed close phylogenic relationships with Nocardioides sp. C157 and Nocardioides simplex. Internal transcribed spacer (ITS) sequences of strain SP12 were longer than those of other Nocardioides sp. and present Ala- and Ile-tRNA unlike Actinomycetales. Nocardioides sp. SP12 presents a novel atrazine catabolic pathway combining trzN with atzB and atzC. Atrazine biodegradation ends in a metabolite that co-eluted in HPLC with cyanuric acid. This metabolite shows an absorption spectrum identical to that of cyanuric acid with a maximal absorption at 214.6 nm. The mass of the atrazine metabolite is in concordance with that of cyanuric acid according to mass spectrometry analysis. Quantitative PCR revealed that the ITS sequence of Nocardioides sp. SP12 was at a lower number than the one of trzN in atrazine-treated soil samples. It suggests that trzN could also be present in other atrazine degrading bacteria. The numbers of trzN and ITS sequences of Nocardioides sp. SP12 were higher in the maize rhizosphere than in bulk soil.

Actinomycetales↗

PubMed2001

Development of a PCR test for the detection of Curtobacterium flaccumfaciens pv. flaccumfaciens.

A chromosomal DNA library of the bacterial pathogen of bean, Curtobacterium flaccumfaciens pv.flaccumfaciens NCPPB 559 was constructed in the plasmid pGEM-7Zf(+). Several clones were identified that hybridised to all Curtobacterium flaccumfaciens pathovars including: C. f betae, C. f flaccumfaciens, C. f oortii, C. f. poinsettiae and, in addition, to some strains of Clavibacter michiganensis subsp. insidiosus and Clavibacter michiganensis subsp. One of these clones (pPMP-26), after subsequent digestion with restriction endonucleases EcoRI/SacI, yielded a fragment of approximately 0.2 Kb (pPMP-26D) that hybridised specifically to C. f flaccumfaciens and not to any of the other plant pathogenic members of the order Actinomycetales or any of the other prokaryotic bean pathogens tested. This fragment was subcloned and sequenced, analysis of the resultant 198 bp sequence showed that no significant homology existed with any other sequence currently deposited in public databases. Further analysis of these data facilitated the design of PCR primers which were subsequently tested against a wide range of plant pathogenic actinomycetes and other prokaryotic bean pathogens. Results show that these primers are highly specific for all strains of C. f flaccumfaciens with no cross-reaction to strains from any other bacterial taxa tested.

Actinomycetales↗

PubMed2000

Artificial chromosomes for antibiotic-producing actinomycetes.

Bacteria belonging to the order Actinomycetales produce most microbial metabolites thus far described, several of which have found applications in medicine and agriculture. However, most strains were discovered by their ability to produce a given molecule and are, therefore, poorly characterized physiologically and genetically. Thus, methodologies for genetic manipulation of actinomycetes are not available and efficient tools have been developed for just a few strains. This constitutes a serious limitation to applying molecular genetics approaches to strain development and structural manipulation of microbial metabolites. To overcome this hurdle, we have developed bacterial artificial chromosomes (BAC) that can be shuttled among Escherichia coli, where they replicate autonomously, and a suitable Streptomyces host, where they integrate site-specifically into the chromosome. The existence of gene clusters and of genetically amenable host strains, such as Streptomyces coelicolor or Streptomyces lividans, makes this a sensible approach. We report here that 100 kb segments of actinomycete DNA can be cloned into these vectors and introduced into genetically accessible S. lividans, where they are stably maintained in integrated form in its chromosome.

Actinomycetales↗

PubMed1969

The configuration of 2,6-diamino-3-hydroxypimelic acid in microbial cell walls.

beta-Hydroxydiaminopimelic acid, together with some diaminopimelic acid, occurs in the cell-wall mucopeptide of certain Actinomycetales. These components were converted into their di-DNP derivatives and separated by chromatography. Hence the relative proportions present in the cell walls of a number of species were measured. The problem of acid-induced inversion of configuration was studied. Of the diaminohydroxypimelic acids isomer B (see Scheme 2; amino groups meso, hydroxy group threo to its neighbouring amino group) always predominated but a small proportion of isomer D (amino groups l, hydroxy group erythro) also occurred. The configuration of the diaminohydroxypimelic acids was determined by periodate oxidation to glutamic gamma-semialdehyde, which underwent spontaneous ring-closure. Reduction with sodium borohydride produced optically active proline, the configuration of which was determined by direct measurement of the optical rotation of DNP-proline. Un-cross-linked diaminohydroxypimelic acid in the cell wall was oxidized with periodate in the presence of ammonia. Since the remaining amino group was bound in peptide linkage, ring-closure was prevented and borohydride reduction of the aldehyde-ammonia presumed to be present resulted in the formation of ornithine. The quantity of ornithine was used as a measure of the degree of cross-linking.

Actinomycetales↗

PubMed2003

Microbial characterization of the Mars Odyssey spacecraft and its encapsulation facility.

Microbial characterization of the Mars Odyssey spacecraft and the Kennedy Space Center Spacecraft Assembly and Encapsulation Facility II (SAEF-II) was carried out by both culture-based and molecular methods. The most dominant cultivable microbes were species of Bacillus, with comamonads, microbacteria and actinomycetales also represented. Several spore-forming isolates were resistant to gamma-radiation, UV, H2O2 and desiccation, and one Acinetobacter radioresistens isolate and several Aureobasidium, isolated directly from the spacecraft, survived various conditions. Sequences arising in clone libraries were fairly consistent between the spacecraft and facility; predominant genera included Variovorax, Ralstonia and Aquaspirillum. This study improves our understanding of the microbial community structure, diversity and survival capabilities of microbes in an encapsulation facility and physically associated with colocated spacecraft.

Actinomycetales↗

PubMed1995

Phylogenetic analysis of the genera Cellulomonas, Promicromonospora, and Jonesia and proposal to exclude the genus Jonesia from the family Cellulomonadaceae.

The 16S rRNA gene sequences of eight Cellulomonas species, two Promicromonospora species, and Jonesia denitrificans were determined, and these sequences were compared with the sequences of about 50 representatives of the Arthrobacter line of descent in the order Actinomycetales. We found that in spite of its current assignment to the family Cellulomonadaceae, J. denitrificans branches outside the radiation of this taxon and cannot be considered a member of it. The two Promicromonospora species do not cluster separately from Cellulomonas species and are more closely related to Cellulomonas species than to each other.

Actinomycetales↗

PubMed1996

The genus Nocardiopsis represents a phylogenetically coherent taxon and a distinct actinomycete lineage: proposal of Nocardiopsaceae fam. nov.

The genus Nocardiopsis was shown to be phylogenetically coherent and to represent a distinct lineage within the radiation of the order Actinomycetales. The closest relatives of the genus Nocardiopsis are members of the genera Actinomadura, Thermomonospora, Streptosporangium, and Microtetraspora. The intrageneric structure of the genus Nocardiopsis is shown to consist of a highly related species group containing Nocardiopsis dassonvillei, Nocardiopsis alborubida, and Nocardiopsis antarctica and a second group of less highly related species comprising Nocardiopsis alba subsp. alba, Nocardiopsis alba subsp. prasina, and Nocardiopsis listeri. Nocardiopsis lucentensis occupies a position intermediate between the two species groups. The results of a 16S ribosomal DNA sequence analysis are generally consistent with the available chemotaxonomic, phenotypic, and DNA-DNA hybridization data. The phylogenetic position and the morpho- and chemotaxonomic properties of Nocardiopsis species support the creation of a family for the genus Nocardiopsis, Nocardiopsaceae fam. nov.

Actinomycetales↗

PubMed1999

Williamsia muralis gen. nov., sp. nov., isolated from the indoor environment of a children's day care centre.

The taxonomic status of an actinomycete (MA 140/96T) isolated from indoor building materials of a children's day care centre was studied using the polyphasic approach. The cell morphology was atypical for an actinomycete, electron microscopy revealed a hairy surface, highly unusual for Gram-positive bacteria. The organisms grew at 10-37 degrees C, no growth was visible at 5 degrees C and 45 degrees C in 5 d. The cell wall contained the diamino acid meso-diaminopimelic acid and the sugars arabinose, galactose, mannose and ribose. The phospholipids phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol and diphosphatidylglycerol were detected. The only menaquinone found was MK-9(H2). The fatty acid pattern was composed of palmitic acid (23.6%) palmitoleic acid (16.5%) and another hexadecenoic acid 16:1cis11 (1.4%), oleic acid (29.9%), stearic acid (2.9%) and the 10-methyl-branched tuberculostearic acid (23.3%). A gas-chromatographic analysis of the mycolic acid revealed a carbon-chain length of C50-C56. The G + C was 64.8 mol%. The results of 16S rDNA sequence comparisons revealed that strain MA 140/96T represents a new lineage in the suborder Corynebacterineae of the order Actinomycetales. Therefore, it was concluded that strain MA 140/96T should be assigned to a new genus and species, for which the name Williamsia muralis gen. nov., sp. nov. is proposed. The type strain of the species is MA 140/96T (= DSM 44343T).

Actinomycetales↗

PubMed1999

Ornithinicoccus hortensis gen. nov., sp. nov., a soil actinomycete which contains L-ornithine.

Two Gram-positive coccoid, non-motile bacteria with L-ornithine as diagnostic diamino acid of the peptidoglycan and an interpeptide bridge of L-Orn<--Gly(1,2)<--D-Glu were isolated from a sample of garden soil. The major menaquinone is MK-8(H4). 13-methyl and 12-methyl tetradecanoic acids are the predominant fatty acids. The polar lipids are phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylserine and two unknown phospholipids. Mycolic acids are absent. The DNA base composition is 72 mol% G + C. Recent comparative 16S rDNA studies revealed that strains HKI 0125T and HKI 0131 represent a novel lineage adjacent to the family Intrasporangiaceae of the order Actinomycetales but distinct from the previously described genera of this family. On the basis of the genotypic, chemotaxonomic, morphological and physiological characteristics of these two isolates it is proposed to classify HKI 0125T and HKI 0131 in a new genus and species for which the name Ornithinicoccus hortensis gen. nov., sp. nov. is proposed. The type strain is HKI 0125T (= DSM 12335T).

Actinomycetales↗

PubMed1999

Beutenbergia cavernae gen. nov., sp. nov., an L-lysine-containing actinomycete isolated from a cave.

Two aerobic, Gram-positive bacteria, strains HKI 0122T and HKI 0132, were isolated from a cave. Cells are not acid-fast, non-motile, non-spore-forming and exhibit a rod-coccus growth cycle. The cell wall peptidoglycan contains lysine in position 3 of the peptide subunit and an interpeptide bridge of L-Lys<--L-Glu. The major menaquinone is MK-8(H4), 13-methyl and 12-methyl tetradecanoic acids are the predominating fatty acids. The polar lipids consist of phosphatidylinositol, diphosphatidylglycerol and three unknown phospholipids. Mycolic acids are absent. The DNA base composition is 71 mol% G + C. Phylogenetic analysis revealed that strain HKI 0122T forms a novel taxon among the families and unassigned genera of the suborder Micrococcineae, within the order Actinomycetales. On the basis of the genotypic, chemotaxonomic, morphological and physiological characteristics of these two isolates it is proposed to assign strains HKI 0122T and HKI 0132 to a new genus and species for which the name Beutenbergia cavernae gen. nov., sp. nov. is proposed. The type strain is HKI 0122T (= DSM 12333T).

Actinomycetales↗

PubMed2000

Frigoribacterium faeni gen. nov., sp. nov., a novel psychrophilic genus of the family Microbacteriaceae.

The taxonomic position of five actinobacterial strains isolated from dust, an animal shed, the air inside a museum and soil was investigated using a polyphasic approach. The growth characteristics were unusual for actinomycetes. Optimal growth was at temperatures ranging from 2 to 10 degrees C. After small-step adaptation (5 degrees C steps) to higher temperatures, the strains were also able to grow at 20 degrees C. Cell wall analyses revealed that the organisms showed a hitherto undescribed, new group B-type peptidoglycan [type B2beta according to Schleifer & Kandler (1972), but with lysine instead of ornithine]. All strains contained menaquinone MK-9. Mycolic acids were not detected. Diphosphatidylglycerol, phosphatidylglycerol and an unknown glycolipid were detected in the polar lipid extracts. The main fatty acids were 12-methyl-tetradecanoic acid (15:0 anteiso), 12-methyl-tetradecenoic acid (15:1 anteiso), 14-methyl-pentadecanoic acid (16:0 iso) and 14-methyl-hexadecanoic acid (17:0 iso), as well as an unusual compound identified as 1,1-dimethoxy-anteiso-pentadecane (15:0 anteiso-DMA). The G+C content of DNA was approximately 71 mol%. The results of 16S rRNA gene sequence comparisons revealed that the strains represent a new lineage in the suborder Micrococcineae and the family Microbacteriaceae of the order Actinomycetales. On the basis of these results the new genus Frigoribacterium gen. nov. is proposed, harbouring the new species Frigoribacterium faeni sp. nov. (type strain = 801T = DSM 10309T).

Actinomycetales↗

PubMed2001

Ornithinimicrobium humiphilum gen. nov., sp. nov., a novel soil actinomycete with L-ornithine in the peptidoglycan.

A Gram-positive bacterium originating from garden soil was taxonomically studied. Cells are non-motile, non-sporulating, irregular rods and cocci. The cell wall peptidoglycan contains L-ornithine as diagnostic diamino acid and an interpeptide bridge consisting of L-Orn<--L-Ala<--Gly<--D-Asp. The major menaquinone is MK-8(H4). 13-Methyl tetradecanoic acid and 14-methyl pentadecanoic acid are the predominant fatty acids. The polar lipids are phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, three unknown glycolipids and three unknown phospholipids. Mycolic acids are absent. The DNA G+C composition is 70 mol%. The acyl type of the glycan chain of peptidoglycan is acetyl. Glucose is the dominating whole cell sugar; arabinose, rhamnose and xylose are present in traces. Results of 16S rDNA sequence comparisons revealed that strain HKI 0124T represents a novel lineage within the suborder Micrococcineae of the order Actinomycetales adjacent to the recently described genus Ornithinicoccus. On the basis of the clearly pronounced morphological, physiological, chemotaxonomic and phylogenetic differences between strain HKI 0124T and all members of the suborder Micrococcineae, it is proposed to assign strain HKI 0124T to a new genus and species, Ornithinimicrobium humiphilum gen. nov., sp. nov. The type and only strain is HKI 0124T (= DSM 12362T = CIP 106634T).

Actinomycetales↗