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Taxonomic study and morphological differentiation of an actinomycete genus, Kitasatospora.

The history and taxonomic criteria of the actinomycete genus, Kitasatospora, are described. The reasons for a change of the position of the genus in the classification system are given and a restriction length polymorphism (RFLP) method for rapid identification is described. Although the bacterial genus, Kitasatospora, resembles the fungal one, Streptomyces, in its morphology, it is clearly different in its cell-wall composition which includes LL- and meso-DAP, glycine and galactose. Following a change in its taxonomic position as a result of insufficient gene analysis, it was re-established by phylogenetic analysis of the entire 16S rRNA gene. Most actinomycetes produce bioactive compounds but the relationship between antibiotic production and morphology has not been studied in detail. Kitasatospora strains may be used as a model because they grow in submerged culture like those actinomycetes strains which produce bioactive compounds. The authors suggest further studies of the correlation between metabolic and morphological differentiation and the mechanism of the production of submerged spores.

Actinomycetales↗

Actinomycetes colonization of tonsils: a comparative study between patients with and without recurrent tonsillitis.

OBJECTIVE: To determine the prevalence of tonsillar actinomycetes colonization in patients with and without recurrent tonsillitis and to study the association of this condition with recurrent tonsillitis and tonsillar hypertrophy. STUDY DESIGN AND SETTING: A retrospective study of 834 patients who had undergone tonsillectomy for recurrent tonsillitis (group A) and for sleep-disordered breathing without a history of recurrent tonsillitis (group B). RESULTS: The prevalence of tonsillar actinomycetes colonization was higher in patients who had undergone tonsillectomy for sleep-disordered breathing (44.1 per cent) than in patients who had undergone tonsillectomy for recurrent tonsillitis (33.3 per cent). The prevalence did not differ by sex or age of patient, although the occurrence rate was higher in the adult compared with the paediatric population. There was no statistically significant difference between the mean size of the tonsils removed in the two groups, and actinomycetes colonization did not affect tonsil size. Histopathological analysis of resected tonsils did not show active tissue infection. CONCLUSION: The presence of actinomyces does not indicate active disease. We are of the opinion that, although actinomyces colonization is more prevalent in patients with sleep-disordered breathing, it does not contribute to tonsillar hypertrophy nor to recurrent tonsillitis.

Actinobacteria↗

Chromosome topology and genome size of selected actinomycetes species.

Information about the genome organization of actinomycetes species is restricted to a few genera: Corynebacterium, Mycobacterium, Rhodococcus, Saccharopolyspora and Streptomyces. Streptomyces species and Saccharopolyspora erythraea were shown to contain a single linear 8 Mb chromosome. In contrast, the Corynebacterium, Mycobacterium and Rhodococcus species studied were demonstrated to possess a smaller (3 Mb-6.5 Mb) single circular chromosome. To investigate whether linear chromosome topology and genome sizes above 7 Mb are unique features of Streptomyces and S. erythraea we have started to investigate the chromosome topology, the genome size and the status of accessory elements of additional actinomycetes species: Actinoplanes philippinensis, Amycolatopsis orientalis, Micromonospora chalcea, Nocardia asteroides, Rhodococcus opacus and Streptoverticillium abikoense. Our data which are based on PFGE experiments clearly suggest that large genome sizes and chromosome linearity are seen in mycelium forming actinomycetes genera. In addition we have identified large linear plasmids in Nocardia asteroides, Streptoverticillium abikoense and Rhodococcus opacus.

Actinomycetales↗

Genomes of 211 Actinomycete Strains from Diverse Environments.

Actinomycetes are a highly diverse group of microorganisms that have long been recognized as a valuable source of antibiotics and other bioactive metabolites. Recent advances in genome mining have revealed a wealth of previously unexplored silent secondary metabolite biosynthetic gene clusters (smBGCs) in actinomycete genomes, underscoring their untapped bioactivity potential. Here, we present the genome sequences of 211 actinomycete strains isolated from various environmental sources, generated through high-throughput sequencing. The resulting genome assemblies exhibit high completeness and accuracy, offering high-quality data for downstream analyses and biological resource exploration.

Actinobacteria↗

Cloning, expression, and substrate specificity of a fungal chymotrypsin. Evidence for lateral gene transfer from an actinomycete bacterium.

Unlike trypsins, chymotrypsins have not until now been found in fungi. Expressed sequence tag analysis of the deuteromycete Metarhizium anisopliae identified two trypsins (family S1) and a novel chymotrypsin (CHY1). CHY1 resembles actinomycete (bacterial) chymotrypsins (family S2) rather than other eukaryote enzymes (family S1) in being synthesized as a precursor species (374 amino acids, pI/MW: 5.07/38,279) containing a large N-terminal fragment (186 amino acids). Chy1 was expressed in Pichia pastoris yielding an enzyme with a chymotrypsin specificity for branched aliphatic and aromatic C-terminal amino acids. This is predictable as key catalytic residues determining the specificity of Streptomyces griseus chymotrypsins are conserved with CHY1. Mature (secreted) CHY1 (pI/MW: 8.29/18,499) shows closest overall amino acid identity to S. griseus protease C (55%) and clustered with other secreted bacterial S2 chymotrypsins that diverged widely from animal and endocellular bacterial enzymes in phylogenetic trees of the chymotrypsin superfamily. Conversely, actinomycete chymotrypsins are much more closely related to fungal proteases than to other eubacterial sequences. Complete genomes of yeast, gram eubacteria, archaebacteria, and mitochondria do not contain paralogous genes. Expressed sequence tag data bases from other fungi also lack chymotrypsin homologs. In light of this patchy distribution, we conclude that chy1 probably arose by lateral gene transfer from an actinomycete bacterium.

Actinomycetales↗

Antigenic relationships among thermophilic actinomycetes.

Antigens from Micropolyspora faeni, Saccharomonospora viridis, Thermoactinomyces candidus, T. sacchari and T. vulgaris were prepared by growing them on dialysate of trypticase soy broth. Sera from rabbits immunized with these antigens were used to study cross-reactivity between thermophilic actinomycetes by antigen-antibody crossed immunoelectrophoresis and by agar gel double diffusion. Mi. faeni and S. viridis showed some degree of cross-reaction, but both failed to show any cross reactivity with Thermoactinomyces species. Antigens from Thermoactinomyces cross-reacted with members within the genus, but no reactivity against S. viridis or M. faeni antisera was detected. Hence, the presence of antibodies to several thermophilic actinomycetes in the sera of patients may be attributed to the exposure of the individual to different thermophilic actinomycetes rather than to the antigenic cross-reactivity between the organisms.

Animals↗

Degradation of pesticides by actinomycetes.

Actinomycetes have considerable potential for the biotransformation and biodegradation of pesticides. Members of this group of Gram-positive bacteria have been found to degrade pesticides with widely different chemical structures, including organochlorines, s-triazines, triazinones, carbamates, organophosphates, organophosphonates, acetanilides, and sulfonylureas. A limited number of these xenobiotic pesticides can be mineralized by single isolates, but often consortia of bacteria are required for complete degradation. Cometabolism of pesticides is frequently observed within this group of bacteria. When compared with pesticide degradation by Gram-negative bacteria, much less information about molecular mechanisms involved in biotransformations of pesticides by actinomycetes is available. Progress in this area has been seriously hampered by a lack of suitable molecular genetic tools for most representatives of this major group of soil bacteria. Overcoming this constraint would enable a better exploitation of the biodegradation and biotransformation abilities of actinomycetes for applications such as bioremediation and construction of transgenic herbicide-resistant crops.

Actinomyces↗

Comparison of species distribution and antimicrobial susceptibility of aerobic actinomycetes from clinical specimens.

To compare the species distribution and antimicrobial susceptibility of aerobic actinomycetes, we evaluated 366 isolates referred to the Centers for Disease Control from October 1985 through February 1988. We used conventional biochemical tests to identify the various species. Four species accounted for 191 (52%) of aerobic actinomycete isolates: Nocardia asteroides (98 isolates), Actinomadura madurae (42 isolates), Streptomyces griseus (28 isolates), and Nocardia brasiliensis (23 isolates). Sputum and wounds were the most common sources. No isolate was resistant to amikacin, no N. brasiliensis isolate was resistant to sulfamethoxazole or trimethoprim-sulfamethoxazole, and no A. madurae isolate was resistant to ceftriaxone or imipenem. In summary, our findings show that unusual species of aerobic actinomycetes can cause infection, colonization, or both and that antimicrobial resistance varies markedly by species.

Actinomycetales↗

In situ identification of nocardioform actinomycetes in activated sludge using fluorescent rRNA-targeted oligonucleotide probes.

Hitherto, few environmental samples have been investigated by a 'full cycle rRNA analysis'. Here the results of in situ hybridization experiments with specific rRNA-targeted oligonucleotide probes developed on the basis of new sequences derived from a previously described comparative 16S rRNA analysis of nocardioform actinomycetes in activated sludge are reported. Application of the specific probes enabled identification and discrimination of the distinct populations of nocardioform actinomycetes in activated sludge. One of the specific probes (DLP) detected rod-shaped bacteria which were found in 13 of the 16 investigated sludge samples from various wastewater treatment plants, suggesting their importance in the wastewater treatment process. Another probe (GLP2) hybridized with typically branched filaments of nocardioforms mainly found in samples from enhanced biological phosphorus removal plants, suggesting that these bacteria are involved in sludge foaming. The combination of in situ hybridization with fluorescently labelled rRNA-targeted oligonucleotide probes and confocal laser scanning microscopy improved the detection of nocardioform actinomycetes, which often showed only weak signals inside the activated-sludge flocs.

Actinomycetales↗

Antifungal spectra of actinomycetes isolated from tobacco.

Five species (28 strains) of actinomycetes isolated from tobacco were tested for antagonism against 12 species of storage and field fungi associated with tobacco. Two strains of Streptomyces albus were antagonistic against all test fungi. The actinomycetes grew more rapidly, produced more pigment, and had more pronounced antibiotic activity when grown at 36 C than at 28 C. Krasilnikov's synthetic medium, SMK-1, supported the greatest antifungal activity. More of the actinomycetes were antagonistic against more test fungi when grown for 20 days rather than 10 days.

Actinomyces↗

Diversity of fungi, bacteria, and actinomycetes on leaves decomposing in a stream.

Although fungi, bacteria, and specific bacterial taxa, such as the actinomycetes, have been studied extensively in various habitats, few studies have examined them simultaneously, especially on decomposing leaves in streams. In this study, sugar maple and white oak leaves were incubated in a stream in northeastern Ohio for 181 days during which samples were collected at regular intervals. Following DNA extraction, PCR-denaturing gradient gel electrophoresis (DGGE) was performed using fungus-, bacterium-, and actinomycete-specific primers. In addition, fungal and bacterial biomass was estimated. Fungal biomass differed on different days but not between leaves of the two species and was always greater than bacterial biomass. There were significant differences in bacterial biomass through time and between leaf types on some days. Generally, on the basis of DGGE, few differences in community structure were found for different leaf types. However, the ribotype richness of fungi was significantly greater than those of the bacteria and actinomycetes, which were similar to each other. Ribotype richness decreased toward the end of the study for each group except bacteria. Lack of differences between the two leaf types suggests that the microorganisms colonizing the leaf biofilm were primarily generalists that could exploit the resources of the leaves of either species equally well. Thus, we conclude that factors, such as the ecological role of the taxa (generalists versus specialists), stage of decay, and time of exposure, appeared to be more important determinants of microbial community structure than leaf quality.

Acer↗

Occurrence of Rhodococcus coprophilus and associated actinomycetes in feces, sewage, and freshwater.

Freshwater, sewage, and fecal samples from various sources were examined for Rhodococcus coprophilus, associated actinomycetes, Escherichia coli, and fecal streptococci. Rhodococcus coprophilus was isolated consistently from feces of farm animals, poultry reared in proximity to farm animals, freshwater, and wastewater polluted with animal fecal wastes. It was not isolated from samples of human feces. The ratio of R. coprophilus total actinomycetes was higher in feces from cattle, sheep, ducks, and geese than in specimens from pigs, horses, and fowl. In samples from two freshwater streams polluted by fecal material from farm animals, the ratios of R. copropilus to total actinomycetes were similar to those found in fecal specimens from cattle and sheep. Ratios of fecal coliform to fecal streptococci could not distinguish between fresh human and animal fecal samples and, furthermore, were not reflected in the stream waters polluted by animal fecal material. R. coprophilus has potential in water and dairy bacteriology as a specific indicator organism of fecal pollution due to farm animal wastes.

Actinomycetaceae↗

Novel actinomycete isolated from bulking industrial sludge.

A novel actinomycete was the predominant filamentous microorganism in bulking activated sludge in a bench-scale reactor treating coke plant wastewater. The bacterium was isolated and identified as an actinomycete that is biochemically and morphologically similar to Amycolatopsis orientalis; however, a lack of DNA homology excludes true relatedness. At present, the isolate (NRRL B 16216) cannot be assigned to the recognized taxa of actinomycetes.

Journal Article↗

Lignin-solubilizing ability of actinomycetes isolated from termite (Termitidae) gut.

The lignocellulose-degrading abilities of 11 novel actinomycete strains isolated from termite gut were determined and compared with that of the well-characterized actinomycete, Streptomyces viridosporus T7A. Lignocellulose bioconversion was followed by (i) monitoring the degradation of [14C]lignin- and [14C]cellulose-labeled phloem of Abies concolor to 14CO2 and 14C-labeled water-soluble products, (ii) determining lignocellulose, lignin, and carbohydrate losses resulting from growth on a lignocellulose substrate prepared from corn stalks (Zea mays), and (iii) quantifying production of a water-soluble lignin degradation intermediate (acid-precipitable polymeric lignin). The actinomycetes were all Streptomyces strains and could be placed into three groups, including a group of five strains that appear superior to S. viridosporus T7A in lignocellulose-degrading ability, three strains of approximately equal ability, and three strains of lesser ability. Strain A2 was clearly the superior and most effective lignocellulose decomposer of those tested. Of the assays used, total lignocellulose weight loss was most useful in determining overall bioconversion ability but not in identifying the best lignin-solubilizing strains. A screening procedure based on 14CO2 evolution from [14C-lignin]lignocellulose combined with measurement of acid-precipitable polymeric lignin yield was the most effective in identifying lignin-solubilizing strains. For the termite gut strains, the pH of the medium showed no increase after 3 weeks of growth on lignocellulose. This is markedly different from the pattern observed with S. viridosporus T7A, which raises the medium pH considerably. Production of extracellular peroxidases by the 11 strains and S. viridosporus T7A was followed for 5 days in liquid cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Actinomycetales↗

Selection of unusual actinomycetal primary sigma70 factors by plant-colonizing Frankia strains.

Functional adaptations of sigma70 transcriptional factors led to the emergence of several paralogous lineages, each one being specialized for gene transcription under particular growth conditions. Screening of a Frankia strain EaI-12 gene library by sigma70 DNA probing allowed the detection and characterization of a novel actinomycetal primary (housekeeping) sigma70 factor. Phylogenetic analysis positioned this factor in the RpoD cluster of proteobacterial and low-G+C-content gram-positive factors, a cluster previously free of any actinobacterial sequences. sigma70 DNA probing of Frankia total DNA blots and PCR screening detected one or two rpoD-like DNA regions per species. rpoD matched the conserved region in all of the species tested. The other region was found to contain sigA, an alternative primary factor. sigA appeared to be strictly distributed among Frankia species infecting plants by the root hair infection process. Both genes were transcribed by Frankia strain ACN14a grown in liquid cultures. The molecular phylogeny of the sigma70 family determined with Frankia sequences showed that the alternative actinomycetal factors and the essential ones belonged to the same radiation. At least seven distinct paralogous lineages were observed among this radiation, and gene transfers were detected in the HrdB actinomycetal lineage.

Base Sequence↗

Non-Frankia actinomycetes isolated from surface-sterilized roots of Casuarina equisetifolia fix nitrogen.

Based on partial 16S sequences, we previously described a novel group of nonsymbiotic, acetylene reduction activity-positive actinomycetes which were isolated from surface-sterilized roots of Casuarina equisetifolia growing in Mexico. An amplified rRNA restriction analysis confirmed that these actinomycetes are distinct from Frankia, a finding substantiated by a 16S rRNA gene phylogenetic analysis of two of the Mexican isolates. Further support for these actinomycetes being separate from Frankia comes from the very low DNA-DNA homology that was found. Nevertheless, the Mexican isolates may be diazotrophs based not only on their ability to grow in N-free medium and reduce acetylene to ethylene but also on the results from (15)N isotope dilution analysis and the finding that a nifH gene was PCR amplified. A comparison of the nifH sequences from the various isolates showed that they are closely related to nifH from Frankia; the similarity was 84 to 98% depending on the host specificity group. An analysis of complete 16S rRNA gene sequences demonstrated that the two strains analyzed in detail are most closely related to actinobacteria in the Thermomonosporaceae and the Micromonosporaceae.

Actinobacteria↗

Lactose-reversible coaggregation between oral actinomycetes and Streptococcus sanguis.

Freshly isolated strains of oral actinomycetes were obtained from human dental plaque and were tested for the ability to coaggregate with common laboratory stock strains of Streptococcus sanguis. Strains belonging to the genera Actinomyces, Arachnia, Bifidobacterium, and Bacterionema were isolated. Only members of the genus Actinomyces coaggregated with the streptococci, and only Actinomyces viscosus and Actinomyces naeslundii exhibited lactose-reversible interactions. A total of 61 strains, consisting of all of the A. viscosus isolates and 86% of the A. naeslundii isolates, coaggregated; 87% inhibited lactose-reversible coaggregation. On the basis of this property and the altered ability of strains to coaggregate after heat treatment of the cells, we delineated four coaggregation groups. The other 13% of the strains constituted a fifth group, which was characterized by a pattern of closely related interactions that were not reversed by lactose. Compared with previously characterized coaggregation properties determined with stock culture strains of actinomycetes, more than 80% of these fresh isolates exhibited identical coaggregation properties. Thus, most of the coaggregation between freshly isolated oral actinomycetes and streptococci involves lactose-reversible cell-cell interactions, which suggests that such coaggregation is mediated by a network of lectin-carbohydrate interactions between complementary cell surface structures on the two cell types.

Actinomyces↗

Denitrification by actinomycetes and purification of dissimilatory nitrite reductase and azurin from Streptomyces thioluteus.

Many actinomycete strains are able to convert nitrate or nitrite to nitrous oxide (N2O). As a representative of actinomycete denitrification systems, the system of Streptomyces thioluteus was investigated in detail. S. thioluteus attained distinct cell growth upon anaerobic incubation with nitrate or nitrite with concomitant and stoichiometric conversion of nitrate or nitrite to N2O, suggesting that the denitrification acts as anaerobic respiration. Furthermore, a copper-containing, dissimilatory nitrite reductase (CuNir) and its physiological electron donor, azurin, were isolated. This is the first report to show that denitrification generally occurs among actinomycetes.

Ascomycota↗