Abyssomicins-A 20-Year Retrospective View.

Abyssomicins represent a new family of polycyclic macrolactones. The first described compounds of the abyssomicin family were abyssomicin B, C, atrop-C, and D, produced by the marine actinomycete strain Verrucosispora maris AB-18-032, which was isolated from a sediment collected in the Sea of Japan. Among the described abyssomicins, only abyssomicin C and atrop-abyssomicin C show a high antibiotic activity against Gram-positive bacteria, including multi-resistant and vancomycin-resistant strains. The inhibitory activity is caused by a selective inhibition of the enzyme 4-amino-4-deoxychorismate synthase, which catalyzes the transformation of chorismate to para-aminobenzoic acid, an intermediate in the folic acid pathway.

Phytomonospora cypria sp. nov., isolated from soil.

A Gram-stain positive actinobacterial strain, designated KT1403(T), was isolated from a soil sample, collected from Karpaz, Magusa, Northern Cyprus, and characterised using a polyphasic approach. Morphological characteristics and chemotaxonomic data indicated that the strain belongs to the genus Phytomonospora. The cell wall of the novel strain contained meso-diaminopimelic acid and galactose, glucose and mannose as the major sugars in whole cell hydrolysates. The polar lipids in the cell membrane were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, an unidentified aminophospholipid and three unidentified glycolipids. The predominant menaquinones were MK-10(H6) and MK-10(H4). The major fatty acids were found to be iso C15:0 , anteiso C15:0 and anteiso C17:0. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain KT1403(T) belongs to the genus Phytomonospora with a sequence similarity of 99.73 % with Phytomonospora endophytica, the type species of the genus. DNA-DNA hybridization further differentiated strain KT1403(T) from its near phylogenetic neighbour, P. endophytica DSM 45386(T) (29.0 ± 2.2 % DNA relatedness). Therefore, it is proposed that strain KT1403(T) represents a novel species of the genus Phytomonospora, for which the name Phytomonospora cypria sp. nov. is proposed. The type strain is KT1403(T) (=KCTC 29479(T) = DSM 46767(T)).

Developmental cycle and pharmaceutically relevant compounds of Salinispora actinobacteria isolated from Great Barrier Reef marine sponges.

The developmental cycle of the obligate marine antibiotic producer actinobacterium Salinispora arenicola isolated from a Great Barrier Reef marine sponge was investigated in relation to mycelium and spore ultrastructure, synthesis of rifamycin antibiotic compounds, and expression of genes correlated with spore formation and with rifamycin precursor synthesis. The developmental cycle of S. arenicola M413 on solid agar medium was characterized by substrate mycelium growth, change of colony color, and spore formation; spore formation occurred quite early in colony growth but development of black colonies occurred only at late stages, correlated with a change in spore maturity in relation to cell wall layers. Rifamycins were detected throughout the growth cycle, but changed in relative quantity at particular phases in the cycle, with a marked increase after 32 days. Expression of the spore division gene ssgA and the rifK gene for 3-amino-5-hydroxybenzoate synthase responsible for rifamycin precursor synthesis was seen even at early stages of the growth cycle. ssgA expression significantly increased between days 26 and 31, but rifK expression effectively remained constant throughout the growth cycle, consistent with the early synthesis of rifamycin. Factors other than precursor synthesis may be responsible for an observed late increase in rifamycin production. A useful approach for measuring and exploring the regulation of antibiotic synthesis and gene expression in the marine natural product producer S. arenicola has been established.

Characterization of Actinoplanes missouriensis spore flagella.

Actinoplanes missouriensis spores swim with a tuft of flagella. Flagella of newborn spores are wrapped with a membranous sheath. When the sheath is unwrapped, spores start swimming. Flagellar length is kept short, at around 1.9 µm, which covers half the circumference of the spore.

Pseudosporangium ferrugineum gen. nov., sp. nov., a new member of the family Micromonosporaceae.

An actinomycete strain 3-44-a(19)(T) was isolated from sandy soil collected in Bangladesh. The strain formed irregular pseudosporangia directly from aggregated spore chains above the rudimentary aerial mycelium. The pseudosporangia developed singly. Each pseudosporangium contained many small, non-motile, spherical, smooth-surfaced spores in chains. Strain 3-44-a(19)(T) contained meso- and 3-hydroxydiaminopimelic acid in the cell wall and MK-9(H(6)) as the major menaquinone and arabinose, galactose, glucose, mannose, ribose and xylose were present in the whole-cell hydrolysate. The diagnostic phospholipid was phosphatidylethanolamine and iso-C(15 : 0) (24.6 %), C(18 : 1)omega9c (15.5 %), C(16 : 0) (10.6 %), C(18 : 0) (9.4 %), iso-C(16 : 0) (8.6 %) and anteiso-C(15 : 0) (6.0 %) were detected as the major cellular fatty acids. The acyl type of the peptidoglycan was glycolyl and mycolic acids were not detected. The G+C content of the DNA was 73.6 mol%. The chemotaxonomic data indicated that the strain belonged to the family Micromonosporaceae. Phylogenetic analysis based on 16S rRNA gene sequence data also suggested that strain 3-44-a(19)(T) fell within the family Micromonosporaceae. On the basis of phylogenetic analysis and characteristic patterns of 16S rRNA gene signature nucleotides as well as morphological and chemotaxonomic data, this strain should be classified as a member of a new genus and species, Pseudosporangium ferrugineum gen. nov., sp. nov., in the family Micromonosporaceae. The type strain of Pseudosporangium ferrugineum is 3-44-a(19)(T) (=JCM 14710(T) =MTCC9007(T)).

A new genus of the order Actinomycetales, Virgosporangium gen. nov., with descriptions of Virgosporangium ochraceum sp. nov. and Virgosporangium aurantiacum sp. nov.

Four motile spored strains were isolated from soil samples collected in Japan. The cultures formed long, narrow sporangia on short sporangiophores directly on the substrate mycelium. The sporangia develop singly or in clusters above the surface of the substrate. Each sporangium contains a single row of six or more motile spores. Glutamic acid, glucosamine, glycine, alanine and 3-OH-diaminopimelic acid are present in the cell wall; the whole-cell sugars are 3-O-methylmannose, rhamnose, mannose, arabinose, galactose, xylose and glucose; and the predominant menaquinones are 10(H4), 10(H6) and 10(H8). The diagnostic phospholipid is phosphatidylethanolamine. The acyl type of the muramic acid is glycolyl. The G+C content is 71 mol%. Mycolic acids are absent. The chemotaxonomic data indicate that these strains belong to the family Micromonosporaceae. Analysis of 165 rDNA sequences suggested that these organisms fall into a distinct clade within the family Micromonosporaceae for which a new genus, Virgosporangium gen. nov., is proposed containing the species Virgosporangium ochraceum sp. nov. (strains YU655-43T, YU793-41 and YU794-41) and Virgosporangium aurantiacum sp. nov. (strain YU438-5T).

Two bacteria causing farmer's lung: fine structure of Thermoactinomyces vulgaris and Saccharopolyspora rectivirgula.

The fine structure of Thermoactinomyces vulgaris and Saccharopolyspora rectivirgula is described by transmission electron microscopy. These two bacteria are the most common microbes causing farmer's lung. The fine structure of hyphae, germination of endospores and the details of conidial wall layers of T. vulgaris, as well as the fine structure of septate hypha and globose, polygonal conidia of S. rectivirgula are described. The conidial wall of T. vulgaris consisted of an inner multilayered spore coat, intermediate spore coat and outer spore coat. The findings are important for the investigations to find fragments of these bacteria in the lungs of exposed patients and experimental animals.

Effect of Micropolyspora faeni cells and cell wall fractions on rabbit alveolar macrophages.

The reactivity of alveolar macrophages (AM) to cells and cell wall fractions (CWF) of Micropolyspora faeni was investigated. Exposure of cultured AM to M. faeni and its CWF caused the AM to form clumps or aggregates which remained attached to the culture dish surface. Other gram-positive and gram-negative bacteria as well as yeast, zymosan, latex microspheres, and isolated peptidoglycan from Listeria monocytogenes did not cause this response. The response was independent of species source and antibody content of the serum used in culture. The use of heat-inactivated sera negated the role of complement activation in the aggregation of AM. AM cultures required a period of culture before exposure to cells or CWF for this response to occur. This response was both time and dose dependent. Rabbit peritoneal macrophages also exhibited the clumping response. Degradation of a purified CWF, fraction 3, with lysozyme greatly diminished the clumping response. Chemical purification of fraction 3 with periodate, formamide, or trichloracetic acid also decreased this activity. These data suggest that the major active component causing this response is peptidoglycan but that other materials associated with the cell wall may also be important. A soluble-factor chemotactic for normal rabbit AM was found in the culture fluid of AM exposed to fraction 3. M. faeni cells and CWF also caused normal rabbit AM to chemiluminesce.

Autolysis of Thermoactinomyces vulgaris spores lacking carbon dioxide during germination.

Ultrathin sections of early germinating endospores of Thermoactinomyces vulgaris were studied by electron microscope. Only spores aerated with an air-CO2 mixture (5% CO2) grow out, while spores aerated with air (0.03% CO2) lyse by the 25th min of inoculation. The lysis is due to progressive, unlimited degradation of the spore integuments and a lack of cell wall formation around the spore protoplast. The requirement of CO2 for outgrowth could not be replaced by oxaloacetate. CO2 seems to be needed to energize the dormant cytoplasmic membrane of the spore to render it capable of initiating active transport processes and of synthesizing the germ cell wall.

Antigen release from Thermoactinomyces vulgaris by lysozyme treatment.

Lysozyme treatment was used to release antigenic material from Thermoactinomyces vulgaris, one of the microbes associated with farmer's lung. Lysozyme caused degradation of the murein layer visualized as changes and disappearance of the bacterial morphology in scanning electron microscopy. Enrichment of different antigenic components in the lysozyme extract and in the cell residue, respectively, was detected by immunoprecipitation. When lysozyme extract and cell residue antigens were used in enzyme-linked immunosorbent assay to test sera of farmer's lung patients and control persons, it became evident that there was no significant difference between the reactions against the two antigens. However, a number of sera reacted preferentially against one or the other of the two antigens.

[Microbial sources of antigens in the environment of farmer's lung patients (author's transl)]. / Mikrobielle antigenquellen in der Umgebung von Patienten mit farmerlunge.

The actinomycetes Micropolyspora faeni and Thermoactinomyces vulgaris are important sources of antigens in extrinsic allergic alveolitis, the deuteromycete Aspergillus fumigatus is it to a lesser extent. These microorganisms have been sought in hay samples obtained from farmers who on the one hand show farmer's lung symptoms and on the other antibodies to these microorganisms. A relatively good correlation has been found between the immunological and microbiological findings. The conditions necessary for the occurrence of antigen sources in farmer's lung disease are discussed, and the need for prophylactic measures is pointed out.

The sporulation process in Thermomonospora fusca as revealed by scanning and transmission electron microscopy.

The sporulation process in the thermophilic actinomycete Thermomonospora fusca was observed by scanning and transmission electron microscopy. As shown by scanning electron microscopy, spores were produced primarily on aerial hyphae and first appeared as bud-like enlargements at the tips of short multibranched sporophores. Young spores were oval to spherical in shape with a smooth surface. As they matured spores enlarged and developed a rough and globular covering, which was quite fragile and easily detached from the spore. This outer layer, as observed by transmission electron microscopy, was thought equivalent to the sheath of other Thermomonospora species. In cross section, mature spores exhibited a thick spore coat underneath the outer globular layer. This spore coat was usually observed as a single layer, but some spores produced a bilayered coat. No multilayered spore coat or spore cortex was observed in the heat-sensitive spores of T. fusca. They were, therefore, shown to be aleuriospores (microcondia), and not endospores.

Existence of a surface configuration on the aerial spore and aerial mycelium of Micropolyspora.

We report the existence of a surface configuration in a true spore-forming genus having cell wall constituents of type IV. Micropolyspora angiospora, M. caesia and M. faeni freeze-fractured along the wall surface and only had a surface configuration on their aerial mycelium and aerial spores, with none on the substrate mycelium and substrate spore. The surface configuration of Micropolyspora was distinctly more complicated than that of Nocardia. The aerial spores of M. angiospora were characteristic in that they possessed ridges, two kinds of surface configurations (i.e. rodlets and fibres), and a complex pattern on the surface. Some rodlets of this organism were formed of a two-stranded helix, each strand having a diameter similar to that of a fibre.

[Fine structure of the vegetative and spore-forming hyphae of Micropolyspora fascifera]. / Tonkoe stroenie vegetativinykh i sporuliruiushchikh gif Micropolyspora fascifera

Changes in the fine structure of the hyphae were studied in the course of spore formation by Micropolyspora fascifera. Sporulating hyphae differ from vegetative hyphae by a less dense cytoplasm, a large zone of the nucleoid with distinctly fibrillar structure, and a thin layerless cell wall. Spore formation is accompanied with autolysis of the vegetative hyphae, which consists in vacuolization of the hyphae and appearance of a large number of tubular membrane structures within them. Spores are formed, like in Actinomyces spp., by simultaneous division of the hyphae with septa; the structure of sporulating septa is similar to that of the fragmenting mycelium in Nocardia; the structure of mature spores is similar to that in some Actinomyces spp. Therefore, M. fascifera differs from other Micropolyspora spp. not only by the chemical composition of the cells (the presence of nocardiomicolic acids) but also by their structure. Taxonomic position of the species is discussed.