New species of Actinomadura isolated from soils in Turkmenia and their antagonistic properties

New species, i.e. Actinomadura polychroma and Actinomadura umbrina are described. The cell wall of the cultures contains meso-diaminopimelic acid galactose, glucose and madurose. The former species is characterized by short spore chains in the form of spirals or pseudosporangia, smooth spore surface, white aerial mycelium and colourless, yellowish-brown or blue-green substrate mycelium. The cultures of this species have no antagonistic activity with respect to various test-microbes. The type culture of A. polychroma is designated as INA 2755. A. umbrina is characterized by formation of short spore chains, which are straight, hooked or spiral, often branching, smooth spore surface, white scanty aerial mycelium and brownish or black-brown substrate mycelium and soluble pigment of the same colour. The strains of this species inhibit the growth of some gram-positive bacteria and have no activity against gram-negative organisms. The type culture of A. umbrina is designed as INA 2309.     

Formation by Actinomyces malachitospinus sp. nov. of a physiologically active substance that stimulates zygote formation in Mucorales]

A new species Actinomyces malachitospinus sp. nov. is described. It has spiral catenulate spores with spines, a gray aerial mycelium, and a green colonial mycelium, and contains no soluble pigments (including melanoid pigments). The strain of Act. malachitospinus INMI 217 produces a physiologically active substance that stimulates the formation of zygotes in Phycomyces blakesleeanus.     

Ultrastructure of the mycelium and spores of Actinomadura fastidiosa sp. nov]

A new species of the Actinomadura genus, A. fastidiosa sp. nov., is described. The ultrastructure of the vegetative mycelium and spores of this organism was studied. The vegetative cells have a multilayered cell wall, often consisting of five layers with different thickness and electron density. The spores are similar to the vegetative cells by their inner structure but have a thicker wall.     

New species of actinomycete Streptomyces variegatus sp. nov. synthesizing an antibiotic of the alpha-hydroxyketopentaene group

Stroptomyces strains INA T-511 and INA 3946 were isolated from 2 different soil samples. Both the strains produced an antibiotic of the group of alpha-hydroxyketopentaenes. The antibiotic was active against gram-positive bacteria, yeasts and fungi belonging to Penicillium. In addition, the strains produced a pigment antibiotic of the prodigiosin type and a pigment identified as ferroverdin. The study of the cultural and morphological features and the natural variation of the strains has shown that they belong to a new species, designated as Streptomyces variegatus sp. nov. Sveshnikova et Timuk.     

New species of Actinoplanes cyanea sp. nov. and its antagonistic properties

An actinomycete stain 1569 isolated from a Siberian soil sample is described as a representative of a new species, designated as Actinoplanes cyaneus sp. nov. The actinomycete cell wall contains meso-diaminopimelic acid, arabinose, xylose and a non-identified analogue of diaminopimelic acid. The actinomycete forms spherical sporangia with mobile spores. The aerial mycelium is absent. The isolate produces a soluble blue pigment on synthetic media. The pygment investigation showed that it belonged to the group of celicomycin-actinorodine.     

Actinomadura fibrosa sp. nov. isolated from soil

A new species of the genus Actinomadura which belongs to the Actinomadura madurae group of Goodfellow et al. was isolated from soil collected in Togo, West Africa. Traditional taxonomic methods plus contemporary fatty acid analysis techniques were used to establish the position of this species. Both physiological characteristics and fatty acid composition differentiate this strain from previously described species. This culture produces a new polyether antibiotic. It is characterized by the production of white to pink aerial hyphae on a limited number of media. The aerial hyphae appear asporogenous, forming thick fibers and projections instead of true spores. The reverse side is a distinctive reddish orange. This organism is resistant to 5% NaCl and grows at temperatures between 20 and 45 degrees C. Whole cells contain meso-diaminopimelic acid, galactose, glucose, mannose, madurose, phosphatidylinositol, and diphosphatidylglycerol. The menaquinones detected were MK-9(H6) and minor amounts of MK-9(H8). The name proposed for this new species is Actinomadura fibrosa; the type strain is strain NRRL 18348.     

Majority of Actinomadura clinical isolates from sputa or bronchoalveolar lavage fluid in Japan belongs to the cluster of Actinomadura cremea and Actinomadura nitritigenes, and the description of Actinomadura chibensis sp. nov

In Japan during 1996-2004, 21 actinomycete strains that have madurose as the diagnostic cell-wall sugar and show true branching in their substrate and aerial mycelia were isolated from sputa or bronchoalveolar lavage (BAL) fluid of patients with pulmonary infections or who were suspected of having related infections. Chemotaxonomic studies showed that all the isolates belong to the genus Actinomadura. Among them, six and seven strains were classified respectively into clusters of Actinomadura nitritigenes and Actinomadura cremea based on 16S rDNA analyses because their 16S rDNA similarities to those respective species were greater than 99.5%. To our knowledge, this is first report that strains of above two species were isolated from clinical specimens. Neither Actinomadura madurae nor Actinomadura pelletieri strain was isolated, and one new species, Actinomadura chibensis, was proposed; the remaining seven strains were not assigned into any known species, suggesting the presence of another new Actinomadura species.     

Echinomycin production by Actinomadura sp. INA 654]

An actinomycete strain designated as Actinomadura sp. INA 654 was isolated from a chernozem soil sample in the Voronezh Region by the soil sample treatment with millimetric waves (EHF band). The strain produced an antibiotic complex of 2 components, named A-654-I and A-654-II. Investigation of their physico-chemical properties showed that A-654-I was identical to echinomycin, a heteropeptide lactone of the quinoxaline group with antitumor activity, while A-654-II proved to be likely a new natural compound. Production of echinomycin by a representative of the Actinomadura genus was detected for the first time. Up to now, only representatives of the Streptomyces genus were known to produce echinomycin.     

Isolation of new species of the genus Actinomadura on selective media with streptomycin and bruneomycin]

In the screening programme using selective media with streptomycin (25, 50 and 100 gamma/ml) and bruneomycin (0.5, 1 and 2 gamma/ml) 3 unusual actinomycetous cultures forming short, rarely long chains of straight, hook-like or spiral spores with 1 or rarely 2 turns were isolated. The study of the cell wall composition of the isolates showed that the walls contained meso-diaminopimelic acid and a sugar madurose. On the basis of the morphological structure and the cell wall composition the isolates were classified as belonging to Actinomadura. Comparison of the cultures with the Actinomadura species described in the literature showed that the isolates were new species of the genera, i. e. Am. coerulea sp. nov., Am. cremea sp. nov. and Am. salmonea sp. nov.     

Actinomadura mexicana sp. nov. and Actinomadura meyerii sp. nov., two novel soil sporoactinomycetes

The taxonomic position of two soil isolates, strains A288(T) and A290(T) [provisionally assigned to the genus Actinomadura] was clarified in a polyphasic study. The organisms showed a combination of chemotaxonomic and morphological properties typical of actinomadurae. They also formed distinct phyletic lines in the 16S rRNA Actinomadura gene tree; strain A288(T) was associated with A. nitritigenes whereas strain A290(T) was closely related to a group that consisted of A. citrea, A. coerulea, A. glauciflava, A. luteofluorescens and A. verrucosospora. Strains A288(T) and A290(T) showed key phenotypic features which readily distinguish them from one another and from representatives of related validly described species of Actinomadura. It is proposed that the two organisms be classified as new species of the genus Actinomadura. The names proposed for the new taxa are Actinomadura mexicana (A290(T) = DSM 44485(T) = NRRL B-24203(T)), and Actinomadura meyerii (A288(T) = DSM 44485(T) = NRRL B-24203(T)).     

A new genus of the Actinoplanaceae: Planobispora,gen. nov.

Summary A new species of a new genus of the Actinoplanaceae is described, for which the name Planobispora longispora gen. nov. sp. nov. is proposed. The organism is a typical mesophilic, aerobic actinomycete, producing a filamentous growth which is differentiated into a vegetative and an aerial mycelium. The new organism is characterized by the formation of sporangia only on the aerial mycelium and by containing a longitudinal pair of motile spores.     

Saccharomonospora oceani sp. nov. isolated from marine sediments in Little Andaman, India

Two actinomycete strains, designated YIM M11168^T and YIM M11177, were isolated from marine sediment samples from Little Andaman, Indian Ocean, and their taxonomic position was determined by a polyphasic approach. The two Gram-positive, aerobic strains were observed to produce branched substrate mycelium and aerial hyphae but did not fragment, and no diffusible pigment was produced on the media tested. At maturity, spores were formed singly or in pairs on aerial hyphae and substrate mycelium, and occasionally the single ones were borne on long sporophores. The optimum growth was determined to occur at 28 °C, 0–4 % (w/v) NaCl and pH 7.0–8.0. Whole-cell hydrolysates of both strains contained meso-diaminopimelic acid and the diagnostic sugars were determined to be galactose, glucose and arabinose. Their predominant menaquinone was found to be MK-9(H₄). The polar lipids detected in the two strains were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylinositol, phosphatidylmethylethanolamine, phosphatidylethanolamine and two unknown phosphoglycolipids. The major fatty acids (>10 %) identified were iso-C_16:0, iso-C_16:1 H, iso-C_16:0, C_17:1 ω6c for strain YIM M11168^T, iso-C_16:0 and Summed Feature 3 for strain YIM M11177. The G + C contents of the genomic DNAs of both strains were determined to be 71.4 %. DNA–DNA hybridization relatedness values (78.4 ± 3.7 %) of these two isolates supported the conclusion that they belong to the same species. Based on phylogenetic analysis, phenotypic and genotypic data, it is concluded that the two isolates belong to a novel species of the genus Saccharomonospora of the family Pseudonocardiaceae. The name Saccharomonospora oceani sp. nov. (Type strain YIM M11168^T = DSM 45700^T = JCM 18128^T) is proposed for the novel species.     

Streptoverticillium griseoviridum n. sp., a producer of the candidin-amphotericin B group, antifungal heptaene nonaromatic antibiotic 0185]

An actinomyceteous strain LIA-0185 producing a heptaenic non-aromatic antibiotic of the candidin type was isolated from a soil sample taken in the Georgian SSR under the programme of screening antifungal antibiotics. The taxonomic study of the strain showed that it belonged to the series of viridoflavum and had the following main taxonomic features: the sporophores in the whorls, straight, remote: the aerial mycelium from yellow to dark-olive-grey; the substrate mycelium olive; the soluble pigment absent; the melanine pigment was produced on the peptone medium; the culture formed H2S; assimilated glucose, mannose, inozide and to a lesser extent fructose; did not assimilate arabinose, xylose, sucrose, lactose, ramnose and raffinose. The strain inhibited the growth of yeast and fungi, grampositive bacteria and actinomycetes and produced a complex of non-aromatic heptaenic antibiotics. The actinomycete differed from the other whorl cultures. It was classified as a new species Sv. griseoviridum sp. nov. The antibiotic complex was a mixture of 2 components, i. e. I and II present approximately in equal amounts. Component II was analogous to candidin. Component I was a new original substance.     

Description of <Emphasis Type="Italic">Streptomyces neopeptinius</Emphasis> sp. nov., an actinobacterium with broad spectrum antifungal activities

A streptomycete strain producing broad-spectrum antifungal substances was taxonomically characterized. The strain, designated KNF 2047(T) (= SH-09(T) = KCTC 10586BP(T)), was found to form extensively branching aerial and substrate mycelia, and produce spiny-ornamented spores with loose spiral chains. The whole cell hydrolyzates contained major amount of LL-diaminopimelic acid. The major fatty acids of the phospholipids were saturated and branched fatty acids containing 14~17 carbons, and the major isoprenoid quinones were hexa-and octa-hydrogenated menaquinones with 9 isoprene units. The phylogenetic analysis using the 16S rRNA gene indicated that the strain belongs to the genus Streptomyces but forms an independent phyletic line. These results clearly demonstrate that strain KNF2047(T) forms a new center of taxonomic variation within Streptomyces, for which the name Streptomyces neopeptinius sp. nov. is proposed.     

Actinomadura species as antibiotic producers

Screening of antibiotic-producing cultures among Actinomadura showed that definite species mainly produced antibiotics of the same groups. Thus, carminomycins were produced by all the 4 studied strains of A. carminata, maduramycins were produced by 3 strains of A. rubra, prodigiozines were produced by 3 strains of A. madurae and luzopeptines were produced by 6 strains of A. recticatena. Supposedly, new antibiotics with original spectral characteristics were isolated from 2 strains of A. fulvescens. There was a clear-cut relation of the number of the active strains and their antibiotic productivity in definite media to their species. The liquid nutrient media, such as yeast-sucrose, soya-glucose and soya-glucose with cobalt chloride proved to be the most efficient in the primary screening of antibiotic-producing cultures.     

Biosynthetic activity of cultures of different species of the genus Actinomadura

The ability to synthesize antibiotics and exoproteases was studied in 13 species belonging to the Actinomadura genus. The antibiotic and proteolytic activities were found only in A. fulvescens INA-3321 and A. citrea INA-1849. Glycerol was shown to be the best carbon source for the growth and biosynthetic activity of A. fulvescens. A. citrea had a wider spectrum of carbon sources used for the growth and a higher proteolytic and antibiotic activity. A. citrea exerted the maximal biosynthetic activity and the best growth in a medium with starch and xylose. The effect of different nitrogen sources on the growth and biosynthetic activity of A. citrea was also studied.     

Eight new species of the genus Micromonospora, Micromonospora citrea sp. nov., Micromonospora echinaurantiaca sp. nov., Micromonospora echinofusca sp. nov. Micromonospora fulviviridis sp. nov., Micromonospora inyonensis sp. nov., Micromonospora peucetia sp. nov., Micromonospora sagamiensis sp. nov., and Micromonospora viridifaciens sp. nov

A previous phylogenetic study on type strains of the genus Micromonospora and Micromonospora species bearing non-validly published names has pointed towards the species status of several of latter strains. Subsequent studies on morphological, cultural, chemotaxonomic, metabolic, and genomic properties, and on whole cell mass spectrometric analyses by matrix adsorbed laser desorption/ionization time-of-flight (MALDI-TOF) confirmed the species status, leading to the proposal of eight new Micromonospora species: Micromonospora citrea sp. nov., type strain DSM 43903T, Micromonospora echinaurantiaca sp. nov., type strain DSM 43904T, Micromonospora echinofusca sp. nov., type strain DSM 43913T, Micromonospora fulviviridis sp. nov., type strain DSM 43906T, Micromonospora inyonensis sp. nov., type strain DSM 46123T, Micromonospora peucetia sp. nov., type strain DSM 43363T, Micromonospora sagamiensis sp. nov., type strain DSM 43912T and Micromonospora viridifaciens sp. nov., type strain DSM 43909T.     

Antimicrobial compounds produced by Actinomadura sp. AC104 isolated from an Algerian Saharan soil

During a search for nonpolyenic antifungal antibiotics, an actinomycete designated AC104 was isolated from a Saharan soil sample by a dilution agar plating method using a chitin - vitamins B medium supplemented with rifampicin. Isolate AC104 presented the morphological and the chemical characteristics of the genus Actinomadura. On the basis of 76 physiological tests and 16S rDNA analysis, this isolate was determined to be quite different from the known species of Actinomadura. It is active against filamentous fungi and both Gram-positive and Gram-negative bacteria. The production of antibiotic substances was investigated using several culture media. The highest antimicrobial activities were obtained on ISP2 medium. The benzenic extract contained five bioactive spots detected on thin layer chromatography plates. Among these antibiotics, a complex called 104A, which showed the more interesting antifungal activity, was selected and purified by reverse-phase high-pressure liquid chromatography. This complex is composed of four compounds. Ultraviolet-visible, infrared, mass, and 1H nuclear magnetic resonance spectroscopy studies showed that these molecules contain an aromatic ring substituted by aliphatic chains. These compounds differ from the known antibiotics produced by Actinomadura species.     

Characterization and comparative studies ofStreptomyces sp. 246 producing a cytostatic antibiotic

The authors submit the results of taxonomic comparative studies of the strainStreptomyces sp. 246, which produces a polypeptide type cytostatic antibiotic. Strain 246 is characterized by tufts of straight sporophores of the “Rectus-Flexibilis” type, smooth spores arranged in chains (over 10 spores in a chain), yellow aerial and substrate mycelium, a negative test for melanin synthesis, utilization of glucose, arabinose, xylose, mannitol, fructose and rhamnose and inability to grow on sucrose, inositol, raffinose and cellulose. The taxonomic characters ofStreptomyces sp. 246 are identical with those of the strainStreptomyces chrysomallus JA 1449-1 and differ manifestly from those ofStreptomyces antibioticus strains (producing actinomycins, antimycin A and oleandomycin), fromStreptomyces cinereoruber ETH 7451 (producing rhodomycin) and from the strainStreptomyces sp. 4127 (producing actinomycin D).     

Polymorphism of a culture of Actinomyces chromogenes var. trienicus, the producer of chromotrienine]

Variation of Actinomyces chromogenes var. trienicus 141-18 MSU, an organism producing trienin was studied under laboratory conditions. Nine stable spontaneous variants were isolated from the population of the initial culture when grown on Gause medium No. 1. The variants varied in differentiation and biosynthetic capacity, including such characteristics as size and form of the colonies, ability for formation of the aerial mycelium and its colour, capacity for sporulation, form of the spore chains and antibiotic production property. In the secondary structures the spores formed only in 6 variants out of 9 isolates. The spore form and spore membrane surface were close in all sporogenic variants, while there were significant differences in the structure of the sporophores. The variants forming the aerial mycelium of the same colour as that of the initial culture did not differ from it also by the nature of the spore chains (spirals with 3--8 turns). The variants with lighter aerial mycelium than that of the initial population formed straight sporophores or spirals with a small number of the turns (1--3). The comparative study of the antimicrobial spectrum of the variants and the component composition of the synthesized antibiotic complex showed that the asporogenic variants and dwarf variant signifcantly differed with respect to their phenotypes from the other cultures and had no antagonistic action. One of the assporogenic variants had only insignificant activity. All the spore forming variants did not differ from the initial culture in the complex of the antibiotics synthesized.