Isolation, Examination and Characterization of Actinomycetes as a Source of Antimicrobial Agents from Libyan Soil

The rapid increase of bacterial resistance strains to multiple antibiotics has become a global public health concern. In the present study, actinomycetes from different districts of Libyan soil were isolated and screened for their inhibitory activity against pathogenic bacteria and fungi. Three hundred soil samples were taken from 77 diverse ecosystems, including deserts, forests, pastureland, and cropland located in different climatic regions in Libya. A total of 164 actinomycetes were obtained. Of 164 isolates, 38 (23.2%) isolates were morphologically and microscopically characterized by spore chain and surface morphology, aerial and substrate mycelia and soluble pigments. The preliminary classification of the isolates illustrates that all isolates belong to the genus Streptomyces. These isolates were further examined for their antagonistic potential against nine pathogenic bacteria and fungi. Out of 38, 11 (28.9%) isolates showed their capability to produce inhibitory substances against at least two tested strains. Among bacterial strains, Staphylococcus aureus was susceptible to almost all eleven isolates (90.9%), while Streptococcus pyogenes was found to be resistant to most selected isolates (18.2%). The isolate, 063 (Wadan soil-desert zone) was the only isolate that exhibited broad spectrum antimicrobial activity against all tested pathogenic microbes, and hence was selected for further study. According to its cultural, morphological, physiological, and biochemical characteristics, the isolate 063 was identified as Streptomyces rochei. The results obtained indicate that the Libyan soil, particularly in extreme environments, could be a potent source of bioactive metabolites with antimicrobial potency against a wide variety of microbial pathogens. Thus, this investigation suggested that exploring new habitats in unexplored and untapped regions of Libya could provide a promising source of biologically active compounds for therapeutic applications.

Determination of Fungicidal Activities of Six Antagonistic Actinomycetes in Soil and Their Preliminary Identification

[ Objective] The paper was to preliminarily identify the species of six actinomyeetes strains, and to determine the fungicidal activities of their fermenta- tion products. E Method ] The species of six aefinomyeetes strains were identified by morphological method, and the fungicidal activities of the fermentation products of six aetinomycetes strains were systematically determined by series of methods including mycellal growth rate inhibition method, spore germination method, potting and field test. []Result] Morphological identification results showed that six strains belonged to Streptornyces. Biological determination results indicated that the in- hibition rates of the fermentation products of six actinomycetes strains with the concentration of 500μg/nd against the mycelial growth of Fusar/um oxysporum f. sp. Vasinfeetum were all greater than 90% ; the inhibition rates against the myeelial growth of Botrytis cinerea Pers. , A/ternar/a a/zernate and Fusarium oxysporum were also greater than 80%. The inhibition rates of the fermentation products of GZ-204 and GZ-331 strains against the spore germination of Bipolaria sorokiniana and Cercospora sorghl were 97. 8% , 98.2%, 99.5% and 94.6%, respectively. Potting test showed that the protection effects of the fermentation products of GZ-204 and GZ-331 strains on wheat powdery mildew（Erysiphe graminis） were 78.8% and 87.1% , and their cure effects were 62.4% and 68.5% , respectively. Field test showed that the control effects of 200 times fermentation liquids of GZ-204 and GZ-331 strains on wheat powdery mildew were 50.5% and 69. 2%, respective- ly. [Condusion] The research provided the reference for the development of new pesticides with actinomyeetes as the resource.

Isolation and Preliminary Identification of Antibacterial Active Substances of Antagonistic Actinomycetes against Peach Crown Gall(Agrobacterium tumefaciens)

[ Objective ] The paper was to isolate and preliminarily identify the antibacterial active substances of antagonistic actinomyeete strain G19 obtained from the soil highly affected by peach crown gall （Agrobacterium tumefaciens）. [ Method] The antibacterial substances of antagonistic actinomycete strain G19 were ex- tracted using protein precipitation method, then isolated and purified using high performance liquid chromatography and medium-pressure preparative chromatogra- phy. Its molecular weight was determined by MALDI-TOFMS method, and the related functional groups were verified through chemical color reaction. [ Result] Seven peptide portions were produced from the antibacterial substances of antagonistic actinomycete strain G19 with the molecular weights of 900 - 1 300 Da after isolation and purification. It could be also inferred that it contained Cys, and carried with H2O and Na＋. Color reaction of functional groups verified that the sub- stance was polypeptide containing glycosyl. [ Conclusion] The result provided basis for the final definition of the structure of antibacterial substances in antagonistic actinomycete strain G19.

Isolation and phylogenetic assignation of actinomycetes in the marine sediments from the Arctic Ocean

Actinomycetes in five marine sediments collected from the Arctic Ocean atdepths of 43 to 3 050 m were cultivated using a variety of media. A total of 61 actinomycetecolonies with substrate mycelia only were observed, and no colonies with aerial mycelia wereobserved under aerobic conditions at 15℃. From these colonies, 28 were selected to representdifferent morphological types. Denaturing gradient gel electrophoresis (DGGE) was used to check thepurity of isolates and select representatives for subsequent sequencing. Phylogentic analyses basedon nearly full-length 16S ribosomal RNA gene (rDNA) sequences indicated that the actinomycetesisolated were accommodated within genus Rhodococcus of family Nocardiaceae, genus Dietzia of familyDietziaceae, genera Janibacter and Terrabacter of family Instrasporangiaceae and genera Kocuria andA nhrobacter of family Micrococcaceae. One of the strains (P27-24) from the deep-sea sediment atdepth of 3 050 m was found to be identical in 16S rDNA sequence(1474/1474) with theradiation-resistant Kocuria rosea ATCC 187~T isolated from air. More than half of the isolatesshowed the similarities ranging from 99.5% to 99.9% in 16S rDNA sequence to dibenzofran-degrading,butyl 2-ethylhexanoate-hydrolysising and nitrile-metabolizing actinomycetes. All the strainsisolated were psychrotolerant bacteria and grew better on the media prepared with natural seawaterthan on the media prepared with deionized water. Three of them (Dietzia sp. P27-10, Rhodococcus sp.S11-3 and Rhodococcus sp.P11-5) had an obligate growth requirement for salt, confirming that thesestrains are indigenous marine actinomycetes.

Isolation and characterization of potential antibiotic producing actinomycetes from water and sediments of Lake Tana,Ethiopia

Objective:To isolate,evaluate and characterize potential antibiotic producing actinomycetes from water and sediments of Lake Tana,Ethiopia.Methods:A total of 31 strains of actinomycetes were isolated and tested against Gram positive and Gram negative bacterial strains by primary screening.In the primary screening.11 promising isolates were identified and subjected to solid state and submerged state fermentation methods to produce crude extracts.The fermented biomass was extracted by organic solvent extraction method and tested against bacterial strains by disc and agar well diffusion methods.The isolates were characterized by using morphological,physiological and biochemical methods.Results:The result obtained from agar well diffusion method was belter than disc diffusion method.The crude extract showed higher inhibition zone against Gram positive bacteria than Cram negative bacteria.One-way analysis of variance confirmed most of the crude extracts were statistically significant at 95%confidence interval.The minimum inhibitor)concentration and minimum bactericidal concentration of crude extracts were 1.65 mg/mL and 3.30 mg/mL against Staphylococcus aureus,and 1.84 mg/mL and 3.80 mg/mL against Escherichia coli respectively.The growth of aerial and substrate mycelium varied in different culture media used.Most of the isolates were able to hydrolysis starch and urea:able to survive at 5%concentration of sodium chloride:optimum temperature for their growth was 30°C.Conclusions:The results of the present study revealed that freshwater actinomycetes of Luke Tana appear to have immense potential as a source ol antibacterial compounds.

Isolation and characterization of marine-derived actinomycetes with cytotoxic activity from the Red Sea coast

Objective: To isolate and evaluate the cytotoxic activity of different actinomycetes species isolated from the Red Sea coast in Sharm el-Sheikh, Egypt.Methods: Forty actinomycetes strains were isolated from different sediments and seawater samples collected from the Red Sea coast in Egypt. Actinomycetes were recognized by morphological and microscopic examinations. Cell viability and cytotoxicity induced by the crude extracts on breast cancer cell lines MDA-MB-231 were assessed using methylene blue assay. The strains with promising cytotoxic activity were identified by sequencing and amplifying the 16 S r RNA genes. The antibacterial activities of the crude extracts were performed using Kirby–Bauer disc diffusion method.Results: The results indicated that five ethyl acetate extracts exhibited cytotoxicity towards breast cancer cell lines MDA-MB-231. The highest cytotoxic activity was found for the ethyl acetate extracts of EGY2 and EGY39. The isolate EGY3 was identified as a new Streptomyces species, while the actinomycete EGY22 was found to be a member of the genus Nocardiopsis sp. The crude extract of the isolate EGY8 showed slightly high antimicrobial activity against different test microorganisms.Conclusions: The results of the present study reveal that marine sediments of the Red Sea are a potent source of novel species of actinomycetes. The isolates may be useful in discovery of novel bioactive compounds and an important step in the development of microbial natural product research.

Clear felling and burning effects on soil nitrogen transforming bacteria and actinomycetes population in Chittagong University campus, Bangladesh

The effect of forests clear felling and associated burning on the population of soil nitrogen transforming bacteria and actinomycetes are reported at three pair sites of Chittagong University campus, Bangla- desh in monsoon tropical climate. Clear felled area or burnt site and 15-21 year mixed plantation of native and exotic species, situated side by side on low hill having Typic Dystrochrepts soil was represented at each pair site. At all the three pair sites, clear felled area or burnt site showed very significantly （p~_0.001） lower population of actinomycetes, Rhizobium, Nitrosomonas, Nitrobacter and ammonifying as well as deni- trifying bacteria compared to their adjacent mixed plantation. From en- vironmental consideration, this finding has implication in managing natural ecosystem.

Diversity and bioactivity of actinomycetes from Signy Is-land terrestrial soils,maritime Antarctic

The Antarctic represents a largely untapped source for isolation of new microorganisms with potential to produce bio- active natural products. Actinomycetes are of special interest among such microorganisms as they are known to produce a large number of natural products, many of which have clinical, pharmaceutical or agricultural applications. We isolated, characterized and classified actinomycetes from soil samples collected from different locations on Signy Island, South Orkney Islands, in the maritime Antarctic. A total of 95 putative actinomyeete strains were isolated from eight soil samples using eight types of selective isolation media. The strains were dereplicated into 16 groups based on morphology and Amplified Ribosomal DNA Restriction Analysis （ARDRA） patterns. Analysis of 16S rRNA gene sequences of representatives from each group showed that streptomy- cetes were the dominant actinomycetes isolated from these soils; however, there were also several strains belonging to diverse and rare genera in the class Actinobacteria, including Demetria, Glaciibacter, Kocuria, Marmoricola, Nakamurella and Tsukamurella. In addition, screening for antibacterial activity and non-ribosomal peptide synthetase genes showed that many of the actinomycete strains have the potential to produce antibacterial compounds.

Distribution of actinomycetes in oil contaminated ultisols of the Niger Delta(Nigeria)

The distribution of actinomycetes in oil contaminated sandy loam ultisols of the Niger Delta region of Nigeria was studied to aid in understanding the effect of hydrocarbons on indigenous microbial population in tropical soils. The average total counts of actinomycetes in all the oil samples analysed was 10 3 cfu/g. Higher counts of actinomycetes were observed during the dry season than during the wet season. The counts of hydrocarbonoclastic actinomycetes correlated positively with the total count of actinomycetes. The actinomycetes were generally restricted to the top soil(0—10 cm soil depth) although a seemingly deeper(down to 40 cm soil depth) distribution was noticed in the dry season. The isolates included oil degrading species of Actinoplanes, Norcadia, Streptomyces and Streptosporangium. Their high oil utilization ability indicates their positive potential and role in the bioremediation of oil spilled soils.

Bioprospection of marine actinomycetes: recent advances,challenges and future perspectives

In exploring new sources for economically important products, marine environment draws particular attention because of its remarkable diversity and extreme conditions;it is known to produce metabolic products of great value. It represents untapped source for the discovery of novel secondary metabolites with varying potential such as antibiotic, anti-tumor, antifouling and cytotoxic properties. Marine actinomycetes distributed throughout the marine environment from shallow to deep sea sediments have proved to be a finest source for this discovery. Secondary metabolites derived from marine actinomycetes have proved their worth in industries based on the research on their properties and wide range applications. Spotlight of the review is range of marine based actinomycetes products and significant research in this field. This shows the capability of marine actinomycetes as bioactive metabolite producers. Additionally, the present review addresses some effective and novel approaches of procuring marine microbial compounds utilizing the latest screening strategies of drug discovery from which traditional resources such as marine actinobacteria has decreased due to declining yields. The aim is in the context of promoting fruitful and profitable results in the near future. The recent surfacing of new technologies for bioprospection of marine actinomycetes are very promising, resulting in high quality value added products, and will be de?ning a new era for bioactive compounds with medical and biotechnological applications.

Hyoscyamine-producing marine Actinomycetes from Lagos Lagoon sediment

Objective:To isolate and screen Actinoniycetes from Lagos Lagoon soil sediments for production of bioactive metabolites.Methods:Sediment samples were collected from four different locations of Lagos Lagoon and were dried for 2 weeks after which the Actinoniycetes were isolated by serial dilution using the spread plate method on starch casein and Kuster's agar supplemented with 80 ug/mL cycloheximide to prevent fungal growth.The plates were incubated at 28 C for 1-2 weeks.Isolates were selected based on their colonial characteristics as well as their Gram's reaction and subciiltured using the same media for isolation until pure cultures were obtained and incubated at 28 C for 3 d.Thereafter,they were inoculated into starch casein and Kuster's broth media and incubated for 8 d.The secondary metabolites were screened for antimicrobial activity against the following microorganisms:methicillin resistant Staphylococcus aureus.Staphylococcus aureus ATCC 29213.Escherichia coli ATCC 29522.Pseudomonas aeruginosa ATCC 27853.Candida albicans and Enterocolitis faecal is ATCC 29212.Coagulasenegative staphylococci isolated from HIV patients were also used(Staphylococcus warneri.Staphylococcus xylosus and Staphylococcus epidennidis).The antimicrobial metabolites of the Actinoniycetes isolates were identified using gas chromatography(GC).Results:Crude extracts of isolates showed antimicrobial activity against some of the test organisms.The GC data analysis showed the antibiotic profile of these isolates.Conclusions:Analysis of the crude extracts of the isolates using GC method,revealed the presence of antibiotics including an anticholinergic hyoscyamine among other conclusions.

Caerulomycin A—An Antifungal Compound Isolated from Marine Actinomycetes

Actinomycetes have been prolific sources of novel secondary metabolites with a range of biological activities that may ultimately find application as therapeutic compounds. Hence several drug discovery companies are engaged in isolation of novel bioactive metabolites from these microbial sources. Antibiotics form the major class of such bioactive metabolites and have been widely used for treating infectious diseases. One of the most critical problems in clinical practice is the increase of prevalence of drug resistant strains, especially azole resistance among fungi. Due to this, there is a constant need for development of new antifungal antibiotics having novel scaffolds and/or mechanism of action. In our in-house screening program in the quest of novel and superior antifungal compounds, an actinomycetes strain PM0525875 was isolated from a marine invertebrate. The extracts of this microbe showed potent in-vitro antifungal activity against drug resistant fungal strains. The antifungal active peak from the extract obtained by shake flask fermentation was identified by chromatographic and other analytical techniques during bioactivity guided isolation. Later the fermentation conditions were optimized in 30 L fermentor for the production of sufficient amount antifungal compound for complete structural characterization. Consequently the fermented broth extract was subjected to bioactivity-guided fractionation, to isolate the active principle using different preparative chromatographic techniques followed by its characterization. The active principle was characterized to be Caerulomycin A. Minimum inhibitory concentration (MIC) of the compound was found in the range of 0.39 - 1.56 μg/ml against pathogenic fungal test strains. The phylogenetic analysis of producer strain using 16S rRNA sequence showed closest match with Actinoalloateichus cyanogriseus. Herewith we report the isolation of Caerulomycin A from marine invertebrate-associated Actinoalloteichus sp. using optimized medium and fermentation conditions.

Diversity and Bioactivity of Actinomycetes from Marine Sediments of the Yellow Sea

Among the 116 actinomycetes collected from marine sediments of the Yellow Sea,56 grew slowly and appeared after 2?3 weeks of incubation.Among the 56 strains,only 3 required seawater(SW) for growth,and 21 grew well in the medium prepared with SW rather than distilled water(DW),while the remaining 32 grew well either with SW or with DW.Six representatives with different morphological characteristics,including 1 SW-requiring strain and 5 well-growing with SW strains,were selected for phy-logenetic analysis based on 16S rRNA gene.Two strains belong to Micrococcaceae and Nocardiopsaceae respectively.The other 4 strains belong to the family of Streptomycetaceae.In the analyzed 6 strains,one was related to Nocardiopsis spp.and the other three were related to Streptomyces spp.,representing new taxa.Bioactivity testing of fermentation products from 3 SW-requiring strains and 21 well-growing with SW strains revealed that 17 strains possessed remarkable activities against gram-positive pathogen or/and tumor cells,suggesting that they were prolific resources for natural drug discovery.

Distribution and generic composition of culturable marine actinomycetes from the sediments of Indian continental slope of Bay of Bengal

Actinomycetes population from continental slope sediment of the Bay of Bengal was studied. Samples were collected during two voyages of FORV Sagar Sampada in 2004 （May-June） and 2005 （July） respectively from 11 transects （each transect had ca. 200 m, 500 m, and 1 000 m depth stations）. The physicochemical parameters of overlying water, and sediment samples were also recorded. The actinomycete population ranged from 5.17 to 51.94 CFU/g dry sediment weight and 9.38 to 45.22 CFU/g dry sediment weight during the two cruises respectively. No actinomycete colony was isolated from stations in 1 000 m depth. Two-way analysis of variance showed significant variation among stations （ANOVA two-way, P〈0.05）, but no significance was found between the two cruises （ANOVA two-way, P〈0.05）. Populations in stations in 500 m depth in both cruises were higher than that of 200 m depth stations with statistically insignificant difference （ANOVA two-way, P〉0.05）. Three actinomycetes genera were identified. Streptomyces was found to be the dominating one in both the cruises, followed by Micromonospora, and Actinomyces. The spore of Streptomyces isolates showed the abundance in spiral spore chain. Spore surface was smooth. Multiple regression analysis revealed that the influencing physico-chemical factors were sediment pH, sediment temperature, TOC, porosity, salinity, and pressure. The media used in the present study was prepared with seawater. Thus, they may represent an autochthonous marine flora and deny the theory of land runoff carriage into the sea for adaptation to the salinity of the seawater and sediments.

Inhibition of norsolorinic acid accumulation to Aspergillus parasiticus by marine actinomycetes

Thirty-six strains of marine actinomycetes were isolated from a sample of marine sediment collected from the Yellow Sea and evaluated in terms of their inhibitory activity on the growth of Aspergillus parasiticus and the production of norsolorinic acid using dual culture plate assay and agar diffusion methods.Among them,three strains showed strong antifungal activity and were subsequently identified as Streptomyces sp.by 16 S rRNA gene sequencing analysis.The supernatant from the fermentation of the MA01 strain was extracted sequentially with chloroform and ethyl acetate,and the activities of the extracts were determined by tip culture assay.The assay results show that both extracts inhibited mycelium growth and toxin production,and the inhibitory activities of the extracts increased as their concentrations increased.The results of this study suggest that marine actinomycetes are biologically important for the control of mycotoxins,and that these bacteria could be used as novel biopesticides against mycotoxins.

Anticancer property of sediment actinomycetes against MCF-7 and MDA-MB-231 cell lines

ObjectiveTo investigate the anticancer property of marine sediment actinomycetes against two different breast cancer cell lines.MethodsIn vitro anticancer activity was carried out against breast (MCF-7 and MDA-MB-231) cancer cell lines. Partial sequences of the 16s rRNA gene, phylogenetic tree construction, multiple sequence analysis and secondary structure analysis were also carried out with the actinomycetes isolates.ResultsOf the selected five actinomycete isolates, ACT01 and ACT02 showed the IC50 value with (10.13±0.92) and (22.34±5.82) μg/mL concentrations, respectively for MCF-7 cell line at 48 h, but ACT01 showed the minimum (18.54±2.49 μg/mL) level of IC50 value with MDA-MB-231 cell line. Further, the 16s rRNA partial sequences of ACT01, ACT02, ACT03, ACT04 and ACT05 isolates were also deposited in NCBI data bank with the accession numbers of GQ478246, GQ478247, GQ478248, GQ478249 and GQ478250, respectively. The phylogenetic tree analysis showed that, the isolates of ACT02 and ACT03 were represented in group I and III, respectively, but ACT01 and ACT02 were represented in group II. The multiple sequence alignment of the actinomycete isolates showed that, the maximum identical conserved regions were identified with the nucleotide regions of 125 to 221st base pairs, 65 to 119th base pairs and 55, 48 and 31st base pairs. Secondary structure prediction of the 16s rRNA showed that, the maximum free energy was consumed with ACT03 isolate (-45.4 kkal/mol) and the minimum free energy was consumed with ACT04 isolate (?7.6 kkal/mol).ConclusionsThe actinomycete isolates of ACT01 and ACT02 (GQ478246 and GQ478247) which are isolated from sediment sample can be further used as anticancer agents against breast cancer cell lines.

Screening of Actinomycetes from Medicinal Plant Rhizosphere Soils for Industrial Enzymes and Antimicrobial Activity

This study aimed to investigate the production of some metabolites （i.e., antibiotics, amylases and cellulases） of terrestrial actinomycetes isolated from medicinal plant rhizosphere soils. Initially, the soil samples were collected from Camellia sinensis （L） Okuntze., Peuraria mirifca Airy Shaw Suvatabandhua., Ananus comosus Merr., Elephantopus scaber Linn., Orthosiphon grandiforus Bolding., Jatropha multifda Linn. and Senna siamea. To screen and isolate actinomycetes, the soil samples were pretreated by air-drying and subsequent heat incubation. The bacterial isolates exhibiting actinomycetes features were then randomly screened for their production of amylases, cellulases and antibiotics. It was found that 130 isolates （from 136） could produce amylases; 40 （from 107） produced cellulases; and seven （from 45） exhibited antimicrobial activity. The data of this study were preliminary, and yet demonstrated a rich diversity of rhizo-actinomycetes from medicinal plants. Besides, these organisms could be an untapped source for discovering of biotechnologically useful metabolites.

Characterization and Antimicrobial Susceptibility of Actinomycetes from TB Smear Negative and Retreatment Patients in Nairobi, Kenya

Actinomycetes are opportunistic pathogens in immunosuppressive patients. Pulmonary actinomycetes infections display symptoms that mimic Mycobacteria tuberculosis and can be misdiagnosed and treated as pulmonary TB. Actinomycetes can be co-infection with tuberculosis leading to delayed or inappropriate treatment. This study aimed to identify and determine antimicrobial susceptibility profiles of Actinomycetes from the sputum of TB smear negative and re-treatment patients referred to TB reference facilities in Kenya. Sputum specimens were collected and direct smears stained with Gram’s reagents. Culture was done on Mueller Hinton agar and incubated at 35°C for two weeks. Identification was done using phenotypic and biochemical procedures. Confirmation of the isolates was done using Polymerase Chain Reaction. A total of 52/385 (14%) Actinomycetes were isolated and subjected to antimicrobial susceptibility testing using broth microdilution method to determine the Minimum Inhibitory Concentration. Nine antibiotics were tested which included: Amikacin, Amoxicillin/Clavulanic acid, Ceftriaxone, Ciprofloxacin, Clarithromycin, Linezolid, Doxycycline, Trimethoprim-Sulfamethoxazole and Gentamycin. Staphylococcus aureus (ATCC 25923) was used as a control. Most of the isolates were susceptible to the test antibiotics. However, four isolates showed multidrug resistance to Ceftriaxone and Clarithromycin with resistance of 11.5% and 26.9% respectively. Gentamycin and Ciprofloxacin showed the highest susceptibility of 100% and 98.1% respectively. The findings of this study confirm that Actinomycetes are significant pathogens in TB smear-negative cases. Although most antibiotics were susceptible, resistance to few antibiotics was observed;hence, there is a need for proper screening of TB smear-negative cases to detect infections by Actinomycetes and also conduct the antimicrobial susceptibility test to determine which antibiotic is effective.

Actinomycetes from Soil of Lachung, a Pristine High Altitude Region of Sikkim Himalaya, Their Antimicrobial Potentiality and Production of Industrially Important Enzymes

Investigations on actinomycetes are profoundly significant area of research since they form major resource for bioactive compounds, antimicrobials, anticancer agents, immunosuppressants, and biological control agents. A total of 59 actinomycetes were isolated from the soil sample collected from Domang, Lachung, North Sikkim, Sikkim, India. Out of the total isolates, 26 isolates with unique and distinct characteristic features were selected and analysed for antimicrobial activity as well as extracellular enzyme production. Out of 26 isolates, 17 (66%) isolates exhibited different level of growth inhibition against the test microorganism. 12 (47%) isolates showed antifungal activity and six (23%) isolates showed antibacterial activity. Most of the isolates showed antifungal activity. Isolate RCS260 was found to exhibit maximum growth inhibition (60%) against Colletotrichum gloeosporioides MTCC 8021. Isolate RCS252 showed maximum growth inhibition (67%) against Bacillus subtilis MTCC 441. Out of 26 isolates, 14 (54%) isolates exhibited chitinase activity, 25 (96%) isolates showed cellulase production, 20 (77%) isolates produced amylase enzyme and 17 (65%) isolates showed positive for protease activity. Potential isolate RCS260 has been characterized and identified as Streptomyces vinaceus strain RCS260 while isolate RCS252 was identified as Kitasatospora aburavienis strain RCS252. The antagonistic profile of strain RCS260 highlights its potential as antifungal agent against phytopathogens.

Diversity of Cultivable Actinomycetes in Tropical Rainy Forest of Xishuangbanna, China

In order to obtain much more un-known actinomycetes for discovering new drug lead, one hundred soil samples were collected from five national natural protection areas of tropical rain forests, Mengla, Menglun, Mandian, Xiaomengyang and Guanping, in Xishuangbanna, Yunnan, China. 1652 purified cultures of actinobacteria were isolated from these samples by using 5 media. The 16S rRNA gene sequences of 388 selected strains were analyzed, and the phylogenetic analysis was carried out. 35 genera which belong to 8 orders and 14 families of the Class actinobacteria were identified. It is showed from research results that actinomycete diversity in tropical rain forest of Xishuangbanna is the highest comparing with all areas studied in our laboratories before. Selective isolation methods for un-known actinomycetes from soil samples, including medium and inhibitors are discussed in this paper.