Characterization Of Actinomycetes Research Articles

Isolation and characterization of actinomycetes with in vitro antagonistic activity against Fusarium oxysporum and Meloidogyne incognita from the rhizosphere of rapeseed and mustard

Twelve isolates of actinomycetes were isolated from soilsamples collected from the rhizosphere of organically cultivated mustard and rapeseed in Sikkim, India. The isolates were morphologically distinguished for their shape and colour and microscopically characterized using Gram staining. The effective 10 Gram-positive actinomycetes isolates were examined for their bio-control efficacy against Meloidogyne incognita and Fusarium oxysporum in vitro. The isolates were studied for biochemical and molecular characterization using 16s rRNA sequencing. The effective isolate (AC-S-2) from the in vitro and biochemical studies was designated as Streptomyces hydrogenans.

Screening, molecular identification, population diversity, and antibiotic susceptibility pattern of Actinomycetes species isolated from meat and meat products of slaughterhouses, restaurants, and meat stores of a developing country, Iran.

Actinomycetes can colonize surfaces of tools and equipment and can be transferred to meat and meat products during manufacture, processing, handling, and storage. Moreover, washing the meat does not eliminate the microorganisms; it only spreads them. As a result, these opportunistic pathogens can enter the human body and cause various infections. Therefore, the aim of the current study was to screen, identify, and determine the antibiotic susceptibility of Actinomycetes species from meat and meat products in the Markazi province of Iran. A total of 60 meat and meat product samples, including minced meat, mutton, beef, chicken, hamburgers, and sausages, were collected from slaughterhouses, butchers, and restaurants in the Markazi province of Iran. The samples were analyzed using standard microbiological protocols for the isolation and characterization of Actinomycetes. PCR amplification of hsp65 and 16SrRNA genes and sequence analysis of 16SrRNA were used for genus and species identification. The minimum inhibitory concentrations (MICs) of antimicrobial agents were determined by the broth microdilution method and interpreted according to the CLSI guidelines. A total of 21 (35%) Actinomycetes isolates from 5 genera and 12 species were isolated from 60 samples. The most prevalent Actinomycetes were from the genus Mycobacterium, with six (28.6%) isolates (M. avium complex, M. terrae, M. smegmatis, and M. novocastrense), followed by the genus Rhodococcus with five (23.8%) isolates (R. equi and R. erythropolis), the genus Actinomyces with four (19.1%) isolates (A. ruminicola and A. viscosus), the genus Nocardia with four (19.1%) isolates (N. asiatica, N. seriolae, and N. niigatensis), and the genus Streptomyces with two (9.5%) isolates (S. albus). Chicken and sausage samples had the highest and lowest levels of contamination, with six and one isolates. Respectively, the results of drug susceptibility testing (DST) showed that all isolates were susceptible to Ofloxacin, Amikacin, Ciprofloxacin, and Levofloxacin, whereas all of them were resistant to Doxycycline and Rifampicin. The findings suggest that meat and meat products play an important role as a reservoir for the transmission of Actinomycetes to humans, thus causing life-threatening foodborne diseases such as gastrointestinal and cutaneous disorders. Therefore, it is essential to incorporate basic hygiene measures into the cycle of meat production to ensure food safety.

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

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

Molecular characterization of Nocardiopsis species from Didwana dry salt lake of Rajasthan, India

The genus Nocardiopsis is well known to produce secondary metabolites especially antibacterial bioactive compound. Isolation and characterization of bioactive compounds producing novel isolates from unusual habitats are crucial. The present study was aimed to explore Didwana dry salt lake of Rajasthan state in India for the isolation and characterization of actinomycetes. The isolated actinomycetes isolates were characterized based on culture characteristics, biochemical tests and 16S rRNA gene sequencing. The 16S rRNA gene sequence analysis revealed that all the five isolates inhabiting soil of the said dry salt lake of Didwana, Rajasthan belonged to four species of Nocardiopsis viz., N. synnemataformans, N. potens, N. prasina and N. dassonvillei subsp. albirubida. The molecular identification based on 16S rRNA gene sequences was found accurate and robust. The phylogram generated through multiple sequence alignment of all the test isolates of Nocardiopsis revealed that the isolates aroused from a single branch and validated monophyletic association. The present study is the first report of exploring Nocardiopsis isolates from the dry salt lake. These characterized Nocardiopsis isolates isolated from Didwana dry salt lake habitat are novel stains and can be of significance in the detection and utilization of novel bioactive compounds.

Distribution and Characterization of Actinomycetes in Mangrove Habitats (Red Sea, Egypt) with Special Emphasis on Streptomyces mutabilis M3MT483919

Ten sediment samples were gathered from several geographical locations around mangrove habitat, Red Sea coast, Egypt, during summer 2019. Actinobacteria are widespread in most mangrove soil samples. The average actinomycetes counts in sediment samples were ranged from 4 to 15 CFUg-1, also physico-chemical characters for soil samples were determined. Statistical analysis was applied to assess if the geographical location and physico-chemical characters influenced the communities of actinomycetes. A total of 10 actinomycetes were isolated and characterized physiologically and biochemically. The antimicrobial activities of different actinomycetes isolates were assessed. Isolate M3 was chosen as the most promising isolate with broad antagonistic activity against Bacillus subtilis ATCC 6633, Escherichia coli ATCC 19404, Staphylococcus aureus ATCC6538, Pseudomonas aeruginosa ATCC 9027, and Candida albicans ATCC 10231 with inhibition zones ranged from 12.0 ± 0.9 to 20.0 ± 1.9 mm. Genotypic characterization of isolate M3 was made using 16S rDNA sequence analysis and identified as Streptomyces mutabilis M3 with accession number MT483919. This strain exhibited anticancer activity against breast cancer cell line (Mcf7), liver cancer cell line (HepG2) and colon cancer cell line (HCT116) and the IC50 values were 324.77, 333.71 and 354.46, respectively. Streptomyces mutabilis M3 MT483919 had high bio-flocculating activity for seawater treatment, and the recovery of the samples ranged between 71.97 and 76.05%. The crude extract of Streptomyces mutabilis MT483919 M3 was analyzed by Fourier transform infrared spectrum (FT-IR) and Gas chromatography-mass spectrometry (GC-MS).

Isolation and characterization of actinomycetes from Mural paintings of Snu- Sert-Ankh tomb, their antimicrobial activity, and their biodeterioration

A total of 35 actinomycetes were isolated from the surface of mural paintings of snu- sort- ankh in El Leasht, Egypt during four seasons all over 2016-2017. In 2009 this tomb was deteriorated by "Aspergillus niger, A. flavus, Fusarium moniliforme, Alternaria alternate, Rhizopus stolonifera, Bacillus subtilis, Bacillus cereus, and micrococcus iuteus." In 2017 the isolation of swabs presents only Aspergillus niger and about 35 actinomycetes classified to three different groups "Streptomyces, Nocardia, and Micromonospora" only five species belong to Streptomyces group showed antimicrobial activity against the previous microorganisms. These actinomycetes were identified according to their sequences in the GenBank to "Streptomyces spectabilis, S. alborgriseolus, S. globsus, S. corchorstt, S. ambofactens." In the other hand, the pigments of the wall paintings of the tomb (Egyptian blue, Egyptian green, Goethite) that analyzed by SEM-EDX, FTIR were measured by the spectrophotometer in both 2009 and 2017. The results showed that the actinomycetes could produce extracellular pigments causing a color change of archaeological pigments otherwise it helps in inhibition the growth of the previous microorganisms found in 2009. The optimization factors for increasing the antibiotic production of the Streptomyces were 3% NaCl and temperature between 30:35°c in Alkin pH (pH = 7.5).

Isolation and Characterization of Actinomycetes from Visakhapatnam Coast and Screening for Antimicrobial Activities of Actinomycetes against Pathogens

Actinomycetes are one of the most attractive sources of antibiotics. In the present study, 32 isolates were isolated from marine soil sediments from the Bay of Bengal, Visakhapatnam. Isolation of Actinomycetes strain was obtained by serial dilution method and grown on actinomycetes isolation agar. Isolated strains were identified for their antibacterial activity of which only five isolates showed good result. They were evaluated for their inhibitory activity on 10 strains of microorganisms (Staphylococcus aureus (MTCC (Microbial Type Culture Collection) 3160), Bacillus subtilis (MTCC 441), Pseudomonas aeruginosa (MTCC 424), Bacillus cereus (MTCC 430), Proteus vulgaris (MTCC 426), Escherichia coli (MTCC 443), Saccharomyces cerevisiae (MTCC 170), Aspergillus niger (MTCC 961), Aspergillus flavus (MTCC 3396), Candida albicans (MTCC 227)). Antibacterial compounds were produced by submerged fermentation and their activity was checked against bacterial culture by antibiogram analysis where extracellular compounds showed positive result. Whereas KRBT-205, KRBT-401 and KRBT-411 showed antibiotic activity against all tested bacterial organisms, the zone of inhibition ranged from 10 mm to 24 mm and KRBT-103 and KRBT-401 showed antibiotic activity against all tested fungal organisms, the zone of inhibition ranged from 10 mm to 24 mm 10mm to 26 mm

Antimicrobial Activity of Actinomycetes Isolated from Different soil samples of Vellore Region

The main objective of the present study was isolation, purification, and characterization of actinomycetes from soil samples. Soil samples were taken from different niche habitats of Vellore district, Tamil Nadu, India. Potential isolate was screened morphologically and biochemically characterised, further were subjected to extraction for the creation of the antibacterial compound. The strains JS26, JS27, and JS28 were discovered profoundly potential, Isolate JS27 displayed the most noteworthy activity against Bacillus cereus (22 mm) and Enterococcus faecalis (25 mm). Ethyl acetate extract showed 89% restraint towards free radicals DPPH (1,1-Diphenyl-2-picrylhydrazyl). The gas chromatography-mass spectroscopy (GC-MS) demonstrated the few unstable mixes. Consequently the outcome showed that the ethyl acetate concentrate of confine groups have tendency to prevent cancer and could be utilized as a part of the advancement of new bioactive metabolites for pharmaceutical purposes.

Isolation and characterization of actinomycetes from marine soil

Four different actinomycetes were obtained from the sampling sites. The isolate AA1 isolated from Puducherry beach exhibited good antibacterial activity on primary screening when compared to the others and hence was chosen for further study. The isolate AA1 was Gram positive with smooth surface morphology and the spore chain morphology showed them to be simple rectus. When the isolate was grown on different media the ariel mycelia was mostly white while the substrate was cream/white. It was able to grow at a pH of 7 at 30°C and tolerate 7% NaCl. Maltose, lactose and xylose was used as the sole carbon source. The crude ethyl acetate extract of AA1 showed antibacterial activity against E.coli (9mm), S.aureus (7.5mm), P.mirabilis (7.7mm), P.aeruginosa (7mm) and K.pneumoniae (4.5mm) at a concentration of 3.5mg/ml. MIC of the crude extract of AA1 was 0.531mg/ml for E.coli and P.aeruginosa, 1.062mg/ml for S.aureus, 0.265mg/ml for P.mirabilis, and 2.125mg/ml for K.pneumoniae. The FT-IR Spectrum of the crude extract showed characteristic peaks at 3290.56cm-1, 1635.64cm-1 and 1016.49cm-1 corresponding to the following functional group such as alcohol (O-H), Alkene (C=C), Ether (C-O) respectively. The 16S rRNA gene sequences of the isolate AA1 revealed 100% similarity to Streptomyces lincolnensis. The restriction sites prediction of the 16SrRNA gene of Streptomyces lincolnensis showed restriction sites for Eco53KI, Bsr I, Hae II, Eco RV, EcoRI, EagI, Hinc II etc. The restriction site analysis showed GC content of 59% and AT content of 41%.

Characterization of ACTINOMYCETES Isolated From Rhizosphere of PISUM SATIVUM L.

Characterization of ACTINOMYCETES Isolated From Rhizosphere of PISUM SATIVUM L.

Characterization of Actinomycetes and Trichoderma spp. for cellulase production utilizing crude substrates by response surface methodology.

Laboratory bench scaling was done and an average of 1.85 fold increase by Response Surface Methodology (RSM) optimization was obtained. It was found that the predicted value (4.96 IU/ml) obtained by RSM is in close accordance with observed activity 5.14 IU/ml. Endoglucanases are mainly induced by CMC while Wheat bran (natural substrate) exoglucanase is more active when induced by avicel and cellulose. Addition of substrate beyond a level caused inhibition of cellulase production. The molecular weight of protein as determined by SDS-PAGE is very similar to molecular weight of cellulase of Trichoderma viride (T. viride) cellulase and Trichoderma reesei (T. reesei) endoglucanase. T. reesei β-glucosidase has high enzymatic activity on CMC substrate when compared with T. viride β-glucosidase. Secondary structure analysed by using Circular Dichroism confirmed that composition of celluase system is very similar to other analysed species. The cellulase was found to be active in pH range of 4.8-5.5; while temperature range varied from 50°C to 70°C. Although the enzymatic activity produced by mutants were lesser than the parent, but in one case mutants of Trichoderma reesei’s BGL has shown higher activity on cellulose.

Physicochemical characterization of actinomycetes isolated from decayed cedar wood: contact angle measurement

Physicochemical characterization of microorganism is very important in a wide range of scientific and technological fields. In this study, we reported the isolation and the molecular identification of actinomycetes recovered from cedar wood decay. The isolates named H5 and H8 were identified by 16S rDNA sequencing and were shown to belong to the genus Nocardia and Streptomyces, respectively. Furthermore, physicochemical proprieties including hydrophobicity, electron donor/acceptor, and the Lifshitz–van der Waals (γLW) of these strains were evaluated using contact angle measurements. The results showed that Nocardia sp. (H5) had a hydrophobic (ΔGiwi = −78.56 mJ/m2) and a weak electron donor/acceptor character. In contrast, results from contact angle measurements showed that the surface free energy of Streptomyces strains (H2, H3, and H8) were ΔGiwi = 20.71 mJ/m2, ΔGiwi = 30.63 mJ/m2, and ΔGiwi = 15.35 mJ/m2, respectively, classifying these microorganisms as hydrophilic bacterium. Moreover, the three strains were predominantly electron donating (γ– ) and exhibit a weak electron-accepting (γ+) character.

Isolation and Characterization of Actinomycetes from Acidic Cultures of Ores and Concentrates

Mining industry is a source of wealth but also of environmental pollution in Peru. In this study 12 colonies of actinomycetes were isolated in acidic cultures from mineral ores and concentrates from mines of the Peruvian highlands. The isolates were characterized phenotypically by microscopy and growth at different conditions as pH tolerance, temperature, and sodium chloride, heavy metals resistance; ferrous iron and thiosulfate oxidation. All isolates were identified as actinomycetes based on their cultural and spore characteristics. Most of the isolates were able to grow at 8 - 45°C and pH 4 - 11. 60% of isolates grew at 10% NaCl but none of them growth at 13%. Iron oxidation was shown by 60% of isolates at pH 4, but only 25% were able to oxidize iron at pH 2. Thiosulfate oxidation was not detected in any isolate. Most of the isolates showed capacity to grow in medium with 200 ppm of Pb, Fe, Zn and 100 ppm of Cu. All of the physiological characteristics found in this work indicate the potential of these isolates as source for bioremediation and bioleaching.

Antibacterial activity of actinomycetes isolated from different soil samples of Sheopur (A city of central India)

The main objective of the present study was isolation, purification, and characterization of actinomycetes from soil samples, having antimicrobial activity against 12 selected pathogenic strains. Soils samples were taken from different niche habitats of Sheopur district, Madhya Pradesh, India. These samples were serially diluted and plated on actinomycete isolation agar media. Potential colonies were screened, purified, and stored in glycerol stock. Isolates were morphologically and biochemically characterized. These isolates were subjected to extraction for production of the antibacterial compound. Antibacterial activity and Minimum Inhibitory Concentration (MIC) of the purified extract of isolates were evaluated. Totally 31 actinomycete isolates were tested for antagonistic activity against 12 pathogenic microorganisms. Isolates AS14, AS27, and AS28 were highly active, while AS1 showed less activity against the pathogenic microorganisms. Isolate AS7 exhibited the highest antagonistic activity against Bacillus cereus (24 mm) and AS16 showed the highest activity against Enterococcus faecalis (21 mm). MIC was also determined for actinomycete isolates against all the tested microorganisms. MIC of actinomycete isolates was found to be 2.5 mg/ml against Shigella dysenteriae, Vancomycin-resistant enterococci, and Klebsiella pneumoniae, and was 1.25 mg/ml for Staphylococcus saprophyticus, Streptococcus pyogenes, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus, Bacillus cereus, Staphylococcus xylosus, Methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, and Staphylococcus aureus. All actinomycetes isolates showed antibacterial activity against S. aureus, while they showed less activity against S. dysenteriae. These isolates had antibacterial activity and could be used in the development of new antibiotics for pharmaceutical or agricultural purposes.

Diversity of Streptomyces spp. in Eastern Himalayan region - computational RNomics approach to phylogeny.

Isolation and characterization of actinomycetes from soil samples from altitudinal gradient of North-East India were investigated for computational RNomics based phylogeny. A total of 52 diverse isolates of Streptomyces from the soil samples were isolated on four different media and from these 6 isolates were selected on the basis of cultural characteristics, microscopic and biochemical studies. Sequencing of 16S rDNA of the selected isolates identified them to belong to six different species of Streptomyces. The molecular morphometric and physico-kinetic analysis of 16S rRNA sequences were performed to predict the diversity of the genus. The computational RNomics study revealed the significance of the structural RNA based phylogenetic analysis in a relatively diverse group of Streptomyces.

Isolation and characterization of actinomycetes from healthy goat faeces

To isolate and characterize actinomycetes with probiotic activities from healthy goat faeces. Faecal actinomycetes were isolated by dilution methods and identified by 16S rRNA gene sequence analysis. The hydrolytic enzyme activities were analysed by clear zone formation. The antimicrobial activities and resistance to heavy metals were tested by growth inhibition methods. The isolates belong to a small group of actinobacterial genera, including Streptomyces, Nocardiopsis and Oerskovia. The Oerskovia was the most widely distributed genus among the cultures. The proportion of streptomycete-like strains producing amylase or protease is significantly higher than those of other actinomycetes (P < 0.05). Compared with streptomycete-like strains, a higher proportion of (alpha- or beta-) galactase-producing other actinomycetes was found in goat faeces. More than 50% of streptomycete-like strains showed activities against test fungi. Streptomycetes could tolerate 0.25 mmol l(-1) Cr(2)O(7)(2-), 2 mmol l(-1) Ni(2+); however, other actinomycetes are liable to 40 mmol l(-1) Fe(3+) and 0.25 mmol l(-1) Cr(2)O(7)(2-) and resistant to 5 mmol l(-1) Ni(2+) and 2 mmol l(-1) Cu(2+). The different physiological characteristics of the actinomycetes suggested that the cooperation in the actinomycetes might be involved in their association with goat. Probiotic mixtures based on faecal actinomycetes showed potentials in animal production.

Isolation and characterization of actinomycetes in vellar estuary, Annagkoil, Tamil Nadu

Vellar Estuary was investigated as a source of actinomycetes to screen for production of novel bioactive compounds. The presence of relatively large populations of Streptomyces in Vellar Estuary soil samples indicates that it is an eminently suitable ecosystem for actinomycetes. Actinomycetes counts ranged 12 x 10 4 cfu/g of soil. The actinomycetes isolated from these ecosystems are capable of producing antibiotics that strongly inhibit the growth of Gram positive and Gram-negative bacteria and yeast like fungi. Further cultural and physiological characterization and DNA homology suggest that strain DPTD-5 is identical to Streptomyces bikiniensis.

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.

Isolation and characterization of actinomycetes from Brazilian tropical soils

Actinomycetes have been isolated from three Brazilian tropical soils. The dispersion and differential centrifugation procedure revealed count values 1.5 to 5.0 times greater than those obtained by the conventional dilution plate technique for all soils and media tested. Eighteen strains, promising for biotechnological applications, were submitted to chemotaxonomic procedures and numerical taxonomy for identification. Two were identified as Amycolatopsis orientalis, one as Streptomyces misakiensis, and two tentatively included or associated to S. chromofuscus and S. griseoruber. The others, all belonging to the Streptomyces genus, could not be fitted into any known species, and were arranged by the UPGMA analysis for classification, as an isolated group. This suggests that the actinomycetes in tropical soils may represent a vast unexplored resource for biotechnology.