ISOLATION AND CHARACTERIZATION OF ACTINOMYCETES FROM SOIL OF AD-DAWADMI, SAUDI ARABIA AND SCREENING THEIR ANTIBACTERIAL ACTIVITIES GOUSE BASHA SHEIK 1* , MUAZZAM SHERIFF MAQBUL (original) (raw)
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International Journal of Pharmacy and Pharmaceutical Sciences, 2017
Objective: To isolate and characterize novel actinomycetes and to evaluate their antibacterial activity against drug-resistant pathogenic bacteria Methods: In the present study, 19 soil samples were collected from different localities of Ad-Dawadmi, Saudi Arabia. Actinomycetes were isolated from these samples using serial dilution and plating method on Actinomycetes isolation agar supplemented with nalidixic acid and actidione to inhibit bacteria and fungi. Crude extracts of potential actinomycetes were produced by submerged fermentation. The antimicrobial activity of crude extracts of actinomycetes was tested against different bacteria using the agar well diffusion method. Characterization of the isolates was done by morphological, physiological and biochemical methods. Results: A total of 9 (47%) isolates of actinomycetes were isolated from 19 different soil samples tested. Among them, 4 (44%) isolates confirmed as Streptomyces sp. showed potential antimicrobial activity against one or more test organisms. Crude extracts were made from these 4 actinomycetes isolates(DOM1, DOM3, DP3, DP4)and tested for their antibacterial activities against 4 different clinical bacterial strains (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus). Crude extract from DP3 isolate showed highest antibacterial activity against all the four test organisms (28 mm, 21 mm, 20 mm and 18 mm) respectively and DP4 showed lowest antibacterial activity against all the four test organisms (14 mm, 12 mm, 0 mm, 6 mm) respectively. The highest zone of inhibition was shown by DP3 against Staphylococcus aureus (28 mm) and Escherichia coli was resistant for DP4. Most of the Inhibition zones produced by crude extracts showed significant differences when compared with control, tested against test organisms (P<0.05). Inhibition zones produced by DP3 and DOM1 against Staphylococcus aureus were 28 mm and 23 mm, respectively which were strong active when compared with control Ciprofloxacin (18 mm). Conclusion: Further studies for purification of bioactive metabolites and molecular characterization analysis of isolated Streptomyces sp. are in progress which would be helpful in discovering novel compounds of commercial value.
Journal of Experimental Biology and Agricultural Sciences
For decades, antibiotics are wonder drugs which treat different microbes and human pathogens such as pneumonia, tuberculosis and gonorrhea which are harder and difficult to treat. Due to miss use of antibiotics in agriculture and animal husbandry, antibiotics are becoming less effective and microbes became more resistant. This resistant increased every second, thus this study aimed to produce active antibiotic from soil actinomycetes which might play a highly significant role in medicine. About 15 bacterial isolates were obtained on starch nitrate agar medium from different soil samples. They were screened for antibacterial production against 5 different human pathogens, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterococcus faecalis and Methicillin-resistant Staphylococcus aureus (MRSA). The most active isolate was SD5 which showed the highest inhibition against E. coli, E. faecalis and MRSA was morphologically examined and characterized. Using molecular identification technique, it was reported that isolate SD5 belonging to genus Streptomyces and was similar to Streptomyces geysiriensis with 97% similarity and to Streptomyces sp. JSM147777 with 95% similarity. Maximum production of the antimicrobial agent was determined by measuring the diameter of inhibition zone using starch nitrate broth, prepared at pH 6.5 and incubation temperature at 30°C for 5 days. The antimicrobial agent was extracted by using three different organic solvents (ethyl acetate, n-Butanol and Petroleum ether). The best solvent was ethyl acetate which gave maximum inhibition against E. faecalis, E. coli and Methicillin-resistant S. aureus. In conclusion, actinomycetes especially
In a screening program to study the antimicrobial activities of desert actinomycetes as potential producers of active metabolites, 75 actinomycete strains were isolated from the Egyptian desert habitats and tested. Out of the isolated 75 organisms, 32 (42.67%) showed activity against the used test organisms. The antimicrobial activities of the active desert actinomycete strains were classified into four groups according to their spectrum of activity on different groups of test organisms and it was found that 43.75% of the active isolates have activity against Gram-positive bacteria only, 28.13% have activity against both Gram-positive and Gram-negative bacteria, 15.63% have activity against Gram-positive bacteria and yeast, 12.50% have activity against Gram-positive, Gram-negative bacteria and yeast. The most potent actinomycete strain, designated D332, was selected for further studies including its identification and isolation of its active compound. Strain D332 was identified by studying its morphology, chemotaxonomy, biochemical characteristics and phylogenetic analysis of the 16S rRNA gene sequence. All phenotypic and genotypic characteristics were consistent with the classification of strain D332 to genus Streptomyces where it formed a distinct phyletic line in the Streptomyces 16S rRNA gene tree. On the other hand, the culture broth of strain D332 was extracted with ethyl acetate after fermentation for the production of the active compound then, the crude extract was partially purified by thin layer chromatography using a solvent system composed of heptane: ethyl acetate (3: 2). The results revealed that strain D332 produced one major compound active against Gram-positive and Gramnegative bacteria and yeasts.
The present study was aimed to isolate and investigate actinomycetes having antimicrobial and cytotoxic activities from soil samples of Mirzapur, Rajshahi, Bangladesh. Total 27 bacteria were isolated and initial screening found that Streptomyces species have low to moderate antagonistic property against various pathogenic bacteria and among them EFAI-1was quite interesting. The antimicrobial activity of crude ethyl acetate extract obtained from EFAI-1was determined using broth-dilution method against Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, Shigella sonnei and Salmonella typhi. The crude extract was almost equally active against both Gram-positive and Gramnegative bacteria. The highest zone of inhibition was found 21 mm at a concentration of 100 µg/disc against Salmonella typhi. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the crude extract against the test bacteria were in the range of 15.6-125 µg/ml and 62.5-250µg/ml, respectively. We found that the LC 50 value of the crude extract was only 0.13 µg/ml against brine-shrimp nauplii indicating its potent cytotoxic nature. Our results indicate that this bacterium would be an excellent source of potent antibiotic and anticancer drugs.
Antimicrobial Compound from Streptomyces Isolate Characterized Using HPLC
2012
Need for novel, safe and more efficient antibiotics is a key challenge to the pharmaceutical industry today. The ever increasing knowledge in the area of pathogen`s drug resistance has evoked the discovery of new antibiotics by the screening of microbes. Last few decades has witnessed the production of novel antibiotics from different microorganisms. At present, aerobic Actinomycetes have attracted considerable attention of bacteriologist, geneticist and ecologist because of the production of novel antibiotics. In this research we evaluate the potential of antibiotic production and characterize HPLC (High performance liquid chromatography) analysis pattern of Streptomyces from various semi-arid locations of Jaipur, Rajasthan. Regarding this, five soil samples were collected randomly from three different green cover areas of Jaipur. Then, following the extraction of secondary metabolite, the HPLC analysis was carried out for characterization of various extracts. Considering the coord...
Life Science Journal
In the present study total 43 actinomycetes RMN1-RMN43 were isolated from the collected soil samples. To screen out active metabolites producing actinomycetes against human pathogenic bacteria, 30 Gram positive and negative bacteria were isolated from hospitalized patients. In the primary screening, 21 actinomycetes showed antibacterial potential against one or more pathogenic bacteria. Total 14 actinomycetes, RMN1-14, showed antibacterial activities through agar well diffusion assay. One isolate of actinomycete, RMN5, produced exclusively narrow spectrum bioactive secondary metabolites against E. coli with 13 mm zone of inhibition. In the secondary screening most potent actinomycete, RMN6, produced broad spectrum bioactive metabolites against ATCC bacterial culture and multi antibiotic resistant bacteria such as E. coli ATCC 25922, P. aeroginosa ATCC 27553, and Staphylococci ATCC 25923 activities at the range of 14, 13 and 19 mm zone of inhibition respectively while against isolate...
2011
Purpose: The rapid emergence of drug resistance among pathogenic bacteria, especially multidrug-resistant bacteria, underlines the need to look for new antibiotics. Methods: In the present study, 134 different actinomycetes, isolated from the soil samples collected from different localities of Punjab and Himachal Pradesh, were screened for antimicrobial activity against various test organisms including multidrug-resistant methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli in order to identify potential antibiotic producers. Results: Among these isolates, 51 (38 %) showed antimicrobial activity against one or more test organisms and six exhibited promising broad-spectrum activity against all the tested organisms. The observed cultural, morphological, physiological and biochemical characteristics confirmed that these isolates are species of the genus, Streptomyces. Conclusion: Further studies on the bioactive metabolites from these cultures will be useful for disc...
Antimicrobial activity of Streptomyces spp. ERI26 recovered from Western Ghats of Tamil Nadu
Journal De Mycologie Medicale, 2008
A new actinomycetes isolate ERI-26 was recovered from Nilgiri forest soil of Western Ghats and screened for its antimicrobial activity against pathogenic bacteria and fungi. Cultural characteristics were assessed in different culture media and 16s rRNA pattern strongly suggested that this isolate belonged to Streptomyces. The morphological and physiological characters were recorded. ERI-26 exhibited resistance to streptomycin. ERI-26 had the ability to produce enzymes such as amylase, cellulase and catalase. Maximum biological activity was obtained on Modified Nutrient Glucose Agar (MNGA) medium. ERI-26 showed activity against bacteria such as Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis and fungi such as Candida albicans, Aspergillus niger and Aspergillus flavus. The antibacterial substances were extracted using methanol from MNGA medium in which ERI-26 had grown for six days at 28 8C. The antimicrobial activity was assessed using broth microdilution technique. The lowest Minimum Inhibitory Concentration of methanol fractions of ERI-26 against S. epidermidis was 375 mg/ml and against C. albicans was 500 mg/ml. # 2008 Elsevier Masson SAS. All rights reserved.
Isolation of actinomycetes with antibacterial activity against multi-drug resistant bacteria
Malaysian Journal of Microbiology
Aims: To isolate and characterise actinomycetes from various sources of soil samples (fruit orchard, dipterocarp forest and oil palm plantation) and to screen these isolates for antibacterial activity against multi-drug resistant bacteria. Methodology and results: A total of 158 fast-growing actinomycete isolates with different colony morphology were subjected to primary cross-streak and secondary well diffusion screening. Six isolates (OP1E, OP7A, OP2A-C, MG1A, UT9C1 and UT7E) were found to inhibit at least one of the seven multi-drug resistant (MDR) bacteria. MG1A exhibited the strongest and broadest spectrum of antibacterial activity against 6 MDR bacteria tested. These isolates were identified as Streptomyces species based on 16S rRNA gene sequence analysis. Further morphological and biochemical analysis revealed that MG1A was highly similar to S. griseocarneus (98.36%) whereas OP1E and OP2A-C were similar to S. parvulus (99.93% and 99.51% respectively). Preliminary identification using LCMS/MS and database search revealed that the major compound in the extract of OP2A-C could be dactinomycin (1255.4170 g/mol). Other antibacterial compounds in the extracts remain to be identified. Conclusion, significance and impact of study: Soil actinomycetes with antibacterial activity against MDR bacteria were isolated not only from undisturbed natural soils but cultivated soils. These isolates were characterised, identified and the antibacterial compounds were extracted for further study. The isolates could serve as a potential source for the development of new and sustainable compounds against MDR bacteria.