A chitinolytic actinomycete complex in black soil

A chitinolytic actinomycete complex in chernozem soil has a specific taxonomic composition, which differs from that of the actinomycete complex which is typically isolated on standard nutrient media containing sugars and organic acids as carbon sources. The actinomycete complex that was isolated by using nutrient media with chitin as the source of carbon and nitrogen was dominated by representatives of the genus Streptosporangium, and the actinomycete complex that was isolated by using nutrient media with sugars and organic acids as the carbon sources was dominated by representatives of the genus Streptomyces. The confirmation to the ability of actinomycetes to utilize chitin as a sole source of carbon and nitrogen came from the augmented length and biomass of the mycelium, the increased number and biomass of the actinomycete spores, the production of carbon dioxide, and the accumulation of NH4+ ions in the culture liquid of the actinomycetes that are grown in the nutrient media with chitin.     

Factors influencing odour production by actinomycetes

Odour production by actinomycete (Streptomyces spp.) strains isolated from hypereutrophic natural waters in which muddy odours in fish have occurred, were studied by the ISP (International Streptomyces Project) carbon utilization method. The streptomycete strains were isolated from water, bottom mud and aquatic plants. Nine different carbon sources were used. Odour character was determined by sniffing the cultures. Odour production varied depending on the strain and the carbon source used. Some of the strains produced similar odours in all media regardless of the carbon source. In other strains, the odour varied depending on the carbohydrate used. The total colony counts of actinomycetes may not necessarily indicate the role of actinomycetes in odour problems in the aquatic environment because the odour production by actinomycetes depends on environmental factors.     

Cultural, morphological, and physiological characteristics of Thermomonospora fusca (strain 190Th).

The cultural, morphological, and physiological properties of Thermomonospora fusca (strain 190Th) are described. Its physiological properties show that this species is primarily a carbohydrate-degrading actinomycete which can use a wide range of plant sugars and polymeric carbohydrates as sources of carbon and energy. The culture does not use proteins or amino acids for carbon and energy, or as a nitrogen source. A few organic acids are utilized. Ammonia is the preferred nitrogen source. The culture has trace nutrient requirements which include biotin and an undetermined number of amino acids. These and other physiological characteristics are discussed in relation to the roles that T. fusca carries out as a saprophytic bacterium in nature. Its cultural and morphological properties are discussed in relation to the taxonomic status of this species in the literature.     

Some Approaches to the Selective Isolation of Actinomycetes of the Genus Actinomadura from Soil

Some approaches to the selective isolation of actinomycetes of the genus Actinomadura from soil are described. The approach that involves the thermal treatment of soil samples and their plating onto Gauze 1 medium with the antibiotics nystatin, nalidixic acid, and rubomycin provides for an increased amount of actinomaduras isolated from the soil actinomycete complex and for a decreased amount of streptomycetes.     

新疆青海中度嗜盐放线菌生物多样性初步研究

从我国新疆、青海等地采集数份盐碱土样或泥样,采用淀粉-酪素琼脂培养基、甘油天门冬酰胺琼脂培养基、土壤浸汁琼脂培养基分别从中分离到8株、32株中度嗜盐放线菌菌株。经形态、生理学特性与全细胞壁氨基酸组分分析结果比较,选取其中的14株进行16S rDNA序列分析。就物种多样性而言,新疆、青海分离到中度嗜盐放线菌分布至少有3个科,5个属。其中有拟诺卡氏菌科（Nocardiopsaceae）的拟诺卡氏菌属(Nocardiopsis)和链单孢菌属（Streptomonospora）;假诺卡氏菌科(Pseudonocardiaceae)的普氏菌属(Prauserella)和糖单孢菌属(Saccharomonospora);链霉菌科(Streptomycetaceae)的链霉菌属(Streptomyces)。就地区分布来讲,新疆分离到的中度嗜盐放线菌种类要远高于青海。     

The Biodiversity of Actinomycetes in Lake Baikal

The taxonomic analysis of 107 actinomycete strains isolated from the bottom sediments and water of Lake Baikal showed that most of the water isolates belong to the genus Streptomyces and most of the sediment isolates belong to the genus Micromonospora. In the sediments, the number of actinomycetes increased with depth (down to 200 m). Eight Streptomyces isolates were identified to a species level.     

Thermophilic chitinolytic microorganisms of brown semidesert soil

In brown semidesert soil, thermophilic prokaryotic organisms identified as Streptomyces roseolilacinus and Silanimonas lenta were shown to play the main role in chitin transformation at 50°C. The phylogenetic positions of the isolated dominant chitinolytic microorganisms were determined on the basis of 16S rRNA gene sequencing. The consumption of chitin as a source of carbon and nitrogen by both the bacterium and the actinomycete was evident from considerable biomass accumulation, high emission of carbon dioxide, and presence in the medium of the chitinase exoenzyme.     

Characterization of actinomycetes isolated from the indoor air of the church of Saint Katherine Monastery, Egypt

Actinomycetes were isolated from the indoor air of the church of Saint Katherin Monastery during different visiting hours. Fifty air samples collected over one year by using automated air sampler were plated on four different media. The low nutrient-content medium 1/10 SC was significantly effective in recovery of actinomycetes compared to the other formula of rich media. Average counts of bacteria, actinomycetes and fungi were 487, 65 and 90 cfu/m³, respectively. Fifty-six morphologically different actinomycetes isolates were recovered during this study assigned into five different genera, in addition to three unidentified isolates. Tentative identification of the isolates indicated predominance of genus Streptomyces, representing 59% of the isolates. Isolates were screened for resistance to 11 antibiotics, antimicrobial activities towards seven microbial strains, Growth on 12 different carbon source, acid production and pigmentation. About 77% of actinomycete isolates were resistant to the antibiotics with different resistance patterns. 12.5% of the airborne actinomycetes exhibited antinmicrobial activities. The isolates showed wide variation in carbon source usage. Forty percent of the isolates were able to utilize all tested carbon sources while 80% were acid producers. Melanin production was produced by 34% of the isolates. While 45% of the isolates were able to produce other diffusible pigments, the majority of the pigments were brown in color however; yellow, orange and green pigments were produced by a number of isolates. The impact of these activities on the historical objects of the church and the tourist’s health is discussed.     

Organic matter fractions by SP-MAS C NMR and microbial communities involved in the suppression of Fusarium wilt in organic growth media

Fusarium wilt is an economically important disease in carnation and tomato plants. The use of suppressive plant growth media has become an alternative method for plant disease control due to the lack of effective chemical control measures. Plant disease suppressiveness is sustained only in plant growth media with an adequate organic matter (OM) composition. Carbohydrate polymers are the most important sources of carbon nutrient for microbial community in these media, mainly consisting of cellulose and hemicellulose. This determines microbial activity, biomass and selects microbial communities in plant growth media, which are reported factors associated with Fusarium wilt suppressiveness.This work determined OM carbon functional groups using Single Pulse Magic Angle Spinning ¹³C-Nuclear Magnetic Resonance (SP-MAS ¹³C-NMR) in three plant growth media with different suppressiveness levels to Fusarium wilt in two crops, carnation and tomato. We propose that the critical role of OM to sustain naturally occurring suppressiveness in those media is not related with cellulose reserve. This could be explained because cellulose protected by lignin encrustation is not available to microbial degradation, meaning that cellulose availability is critical to sustenance of microorganism-mediated biological control. However, the hemicellulose relative abundance (peak 175 ppm) was associated to Fusarium wilt suppression level in plant growth media studied.Carbon source availability in OM was related to microbial biomass and econutritional group population densities involved in biocontrol. For these composts, Bacillus spp., oligotrophic and cellulolytic actinomycetes, and oligotrophic actinomycetes/oligotrophic bacteria and cellulolytic actinomycetes/cellulolytic bacteria ratios were indicated as microbial populations potentially involved in suppression.     

Application of a method incorporating differential centrifugation for selective isolation of motile actinomycetes in soil and plant litter

The present paper describes a simple enrichment technique which enables rapid and selective isolation of diverse zoosporic actinomycete genera directly from soil and plant litter. This technique, designated the rehydration and centrifugation (RC) method, consists of immersing the air-dried source material in 10 mM phosphate buffer containing 10% soil extract, letting the preparation stand at 30 °C for 90 min, followed by centrifugation of the fluid at 1,500×g for 20 min. Portions of the supernatant containing actinomycete zoospores are plated on the humic acid-vitamin agar which is supplemented with nalidixic acid and trimethoprim as the selective inhibitors for Gram-negative bacteria and bacilli. The phosphate buffer-soil extract solution significantly promoted liberation of motile zoospores from the source material. The centrifugation stage greatly eliminated streptomycetes and other non-motile actinomycetes from the liquid phase, thereby facilitating selective growth of rare, motile actinomycetes on the isolation plates subsequent to inoculation. Ten different soil and leaf-litter samples, taken from fields, forests, and stream banks, were examined. The RC method consistently achieved preferential isolation of motile actinomycetes in all samples, which accounted for 37–86% of the total microbial population recovered. The most frequently isolated motile actinomycetes were Actinoplanes and Dactylosporangium. Strains of Actinokineospora, Catenuloplanes and Kineosporia were also recovered, depending on the nature of the samples examined. Other motile actinomycetes that were occasionally isolated in small numbers included Actinosynnema, Geodermatophilus and Sporichthya.     

Alkaliphilic Soil Actinomycetes

The actinomycete complex of alkaline soils was found to be dominated by alkaliphilic streptomycetes, which showed maximal radial rates of colony growth at pH 8. At pH values of 7 and 10, the growth of these streptomycetes was poor. Alkaliphilic streptomycetes can be morphologically differentiated from other actinomycetes based on their high radial rates of colony growth and increased spore formation in alkaline media as compared to neutral media.     

Isolation of Actinomycetes from Soil Using Extremely High Frequency Radiation

A new method employing extremely high frequencies (EHFs) is proposed for the selective isolation of actinomycetes from soil. The pretreatment of soil suspensions with EHF wavelengths of 5.6 and 7.1 mm led to a nonselective isolation of actinomycetes. At the same time, the irradiation of soil suspensions within wavelength bands of 3.8–5.8 and 8–11.5 mm considerably augmented the total number of isolated actinomycetes and increased the fraction of the isolated rare genera by 2 and 7 times, respectively. The rare actinomycete genera were represented by Actinomadura, Microtetraspora, Nonomuraea, Micromonospora, Amycolatopsis, Pseudonocardia, Saccharotrix, and Streptosporangium.     

西沙群岛海域海洋放线菌的分离及其抗菌活性

分离西沙群岛海域放线菌并研究其抗菌活性。采用干燥、辐射、冷冻及加热处理等11种样品预处理方式和10种培养基对海洋放线菌进行分离,对代表性菌株进行鉴定,并考察分离放线菌生长的海水依赖性。进一步以金黄色葡萄球菌、大肠埃希菌、啤酒酵母和扩展青霉为指示菌考察分离放线菌的抗菌活性。从西沙群岛海域样品中分离获得放线菌383株,其中专性海洋放线菌23株。选定93株代表菌株进行鉴定,93株菌隶属于9个科,11个属。不同培养基对分离放线菌菌株的数量及种类影响显著。6株放线菌对4种指示菌均有抑菌活性,其中4株为专性海洋放线菌,表明海洋环境具有丰富的放线菌资源,这些放线菌特别是专性海洋放线菌有望为新型抗菌物质的发现与开发提供菌种来源。     

Unique Actinomycetes from Marine Caves and Coral Reef Sediments Provide Novel PKS and NRPS Biosynthetic Gene Clusters

In the ever-expanding search for novel bioactive molecules and enzymes, marine actinomycetes have proven to be a productive source. While open reef sediment and sponge-associated actinomycetes have been extensively examined, their marine cave counterparts remain unevaluated. Anchialine cave systems in the Bahamas offered an ideal setting to evaluate the occurrence and variation within sediment-associated actinomycete communities. While in close geographical proximity to open reef environments, these systems provide a specialized environmental niche devoid of light and direct exposure to nutrient input. In the present study, selective isolation techniques and molecular methods were used to test the hypothesis that variable distribution of actinomycetes and secondary metabolite gene clusters occur between open reef and marine cave systems. The results indicated that differences exist within the culturable sediment-associated actinomycete communities between marine caves and open reef systems, with members of the genus Streptomyces dominating cultures from open reef sediments and a more diverse suite of actinomycetes isolated from marine cave sediment samples. Within the cave isolates, members of the proposed genus Solwaraspora were the most represented. Based on PKS- and NRPS-gene-targeted PCR amplification and sequencing, geographic variation in the occurrence of these biosynthetic pathways was also observed. These findings indicate that marine cave systems are a lucrative source in the search for novel secondary metabolite producers with biotechnological applications and that environmental and geographic factors likely affect the occurrence of these biosynthetic pathways.     

Isolation and characterization of a unicellular manganese-oxidizing bacterium from a freshwater lake in northwestern Russia

A unicellular manganese-oxidizing bacterium (strain L7), isolated from Lake Ladoga, is identified as “Siderocapsa” sp. according to its morphology. However, this bacterium belongs to the phylogenetic cluster of Pseudomonas putida. The physiological characteristics (utilization of sugars, polyols, organic acids, and phenolic substrates as carbon and energy sources) also indicate the similarity of strain L7 to representatives of the genus Pseudomonas. The growing culture oxidizes Mn(II); the rate of oxidation depends on the type of added organic substrate. Carbonate requirement for this process indicates mixotrophic metabolism. The relatedness of the isolated bacterium to the representatives of the genus Pseudomonas and their phenotypic similarity provide a basis for considering strain L7 not as “Siderocapsa” sp., but as a new species, Pseudomonas siderocapsa sp. nov., of the P. putida cluster.     

The importance of amino acids as carbon sources for Micromonospora echinospora (ATCC 15837)

AIMS: This study set out to investigate the effect of amino acids on the uptake of glucose by Micromonospora eichinospora (ATCC 15837). METHODS AND RESULTS: The specific rate of glucose uptake was found to be reduced when organic nitrogen components were present in the medium. Radioactive uptake studies revealed that the Km for glucose in this organism was 53 mm, indicating a low affinity for uptake compared with other actinomycete sugar transport systems. Individual amino acids negatively influenced the rate of glucose transport, suggesting a relationship between amino acid metabolism and glucose uptake in this organism. The sugar transport system was found to be an active process being inhibited by ionophores and KCN. CONCLUSIONS: The data suggest a direct link between amino acid metabolism and glucose uptake at the level of sugar transport. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that the uptake of glucose, a major carbon source for many antibiotic fermentations, is significantly reduced in the presence of amino acids. This fact should inform the medium design and feeding regimes of fermentations involving similar actinomycetes.     

Isolation and characterization of a unicellular manganese-oxidizing bacterium from a freshwater lake in northwestern Russia

A unicellular manganese-oxidizing bacterium (strain L7), isolated from Lake Ladoga, is identified as "Siderocapsa" sp. according to its morphology. However, this bacterium belongs to the phylogenetic cluster of Pseudomonas putida. The physiological characteristics (utilization of sugars, polyatomic alcohols, organic acids, and phenolic substrates as carbon and energy sources) also indicate the similarity of strain L7 to representatives of the genus Pseudomonas. The growing culture oxidizes Mn(II); the rate of oxidation depends on the type of added organic substrate. Carbonate requirement for this process indicates mixotrophic metabolism. The relatedness of the isolated bacterium to the representatives of the genus Pseudonomas and their phenotypic similarity provide a basis for considering strain L7 not as "Siderocapsa" sp., but as a new species, Pseudomonas siderocapsa sp. nov., of the P. putida cluster.     

A comparison of carbon/energy and complex nitrogen sources for bacterial sulphate-reduction: potential applications to bioprecipitation of toxic metals as sulphides

Detailed nutrient requirements were determined to maximise efficacy of a sulphate-reducing bacterial mixed culture for biotechnological removal of sulphate, acidity and toxic metals from waste waters. In batch culture, lactate produced the greatest biomass, while ethanol was more effective in stimulating sulphide production and acetate was less effective. The presence of additional bicarbonate and H₂ only marginally stimulated sulphide production. The sulphide output per unit of biomass was greatest using ethanol as substrate. In continuous culture, ethanol and lactate were used directly as efficient substrates for sulphate reduction while acetate yielded only slow growth. Glucose was utilised following fermentation to organic acids and therefore had a deleterious effect on pH. Ethanol was selected as the most efficient substrate due to cost and efficient yield of sulphide. On ethanol, the presence of additional carbon sources had no effect on growth or sulphate reduction in batch culture but the presence of complex nitrogen sources (yeast extract or cornsteep) stimulated both. Cornsteep showed the strongest effect and was also preferred on cost grounds. In continuous culture, cornsteep significantly improved the yield of sulphate reduced per unit of ethanol consumed. These results suggest that the most efficient nutrient regime for bioremediation using sulphate-reducing bacteria required both ethanol as carbon source and cornsteep as a complex nitrogen source.     

A new preferential medium for enumeration and isolation of desert actinomycetes

In order to facilitate the discovery of novel actinomycetes from the Egyptian deserts, which can be useful as new sources for bioactive metabolites, different media for enumeration and isolation of desert actinomycetes have been tested. For this purpose, 30 soil samples from different six sites representing the Western and Eastern deserts of Egypt were collected. The two deserts are considered hyper-arid and the soil characteristics were determined. The media used were glucose–yeast extract agar, soil extract agar and a new minimal medium (MM) containing glucose, yeast extract and mineral salts. The effects of the soil characteristics on the total viable actinomycete counts on the three media were evaluated. The results showed that the highest actinomycete count in samples from five out of six sites was obtained on MM. Also MM was more selective for actinomycetes and significantly decreased the number of fungal colonies and to a lower extent the number of bacterial colonies. Moreover, it supported the development of different and diverse groups of actinomycetes. From the results obtained in this study, MM is a new useful medium for enumeration and selective isolation of actinomycetes from the desert soils.     

Repression of alkaline protease in salt-tolerant alkaliphilic <Emphasis Type="Italic">Streptomyces clavuligerus</Emphasis> strain Mit-1 under the influence of amino acids in minimal medium

A salt-tolerant alkaliphilic actinomycete (strain Mit-1) was isolated from Mithapur (Western Coast, Gujarat, India) and identified as Streptomyces clavuligerus. Based on 16S rRNA gene sequence (EU146061) homology, it was found to be related to Streptomyces sp. (AY641538.1). The strain secreted alkaline protease optimally at 5% NaCl and pH 9 during the early stationary phase and could utilize the amino acids methionine, alanine, leucine, phenylalanine, tyrosine, tryptophan, arginine, asparagine, histidine, and glutamic acid as the sole source of nitrogen. Above their threshold levels, these amino acids caused repression of alkaline protease production. Protease production with methionine (120 U/mL), histidine (140 U/mL), and aspartic acid (118 U/mL) was comparable to that with complex medium (130 U/mL). However, the production increased with an increasing number of different amino acids in the growth medium. Repression of protease production as influenced by the amino acids generated valuable information on enzyme synthesis in actinomycetes, as such data is scarce. Optimization of the conditions for enzyme production by actinomycetes in general, and in haloalkaliphilic actinomycetes in particular, appears to be an attractive proposition for biocatalysis.