RESUMO
INTRODUCTION: The actinomycetes are diversely distributed microorganisms in nature. The geographical diversity of Nepal is suitable for adaptation of various species of actinomycetes. The distribution of the actinomycetes is dependent upon the altitude and nature of the soil and water. METHODS: A total of 22 water and soil samples were collected from different regions of Nepal and were processed. Different isolates were characterized by observing colony characteristics and microscopic characteristics. Screening of the antimicrobial property was based upon perpendicular line streaking and submerged-state fermentation for antibiotic production. RESULTS: From the identification tool used, 12 were found to be Micromonospora, 9 were Nocardia, and 7 were Streptomyces. Out of total samples, 8 isolates of actinomycetes were tested effective against the tested bacteria by primary screening using the well diffusion method. Among the primarily screened, all isolates were subjected to submerged-state fermentation methods to produce crude extracts and 4 were found to be effective against the tested bacterial group. The actinomycetes identified from a water source showed better antimicrobial property towards the tested bacteria than the soil sample. Most isolates were found to be Micromonospora followed by Nocardia and Streptomyces with higher antimicrobial activities. CONCLUSION: The water source and soil sediments of Nepal provide suitable environments for actinomycetes towards obtaining a novel antimicrobial agent. The study of actinomycetes from various unexploited areas of Nepal is necessary. Thus, exploitation of various regions of Nepal for the discovery of an effective antimicrobial agent is helpful in providing a solution to the cost-effective therapy and action against antibiotic resistance.
RESUMO
Actinomycetes are Gram-positive, facultative anaerobic fungus-like filamentous bacteria which remain on the top of the natural antibiotic producers. Due to the climatic and geographical diversity of Nepal, a wide range of microorganisms with potent source of antimicrobials are available. The objective of this study was to isolate, identify, and screen the potential antimicrobial-producing actinomycetes from soils covering different altitude range of Nepal. Forty-one isolates of actinomycetes were isolated from 11 soil samples collected from different locations in Nepal with altitude ranging from 1500 to 4380 meters. The isolates were identified on the basis of morphological study, different sugar utilization, protein utilization, and hydrolysis tests. They were also characterized on the basis of temperature and pH. Primary screening for antimicrobial activity was carried out against several test organisms: Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), Klebsiella pneumoniae (ATCC 700603), and Pseudomonas aeruginosa (ATCC 27853) by the perpendicular streaking method, and secondary screening was carried out by the agar well diffusion method using ethyl acetate for solvent extraction. 70.7% of the isolates were identified as Streptomyces spp., 19.5% as Nocardia spp., and 9.5% as Micromonospora spp. 43.34% of actinomycete isolates was found to be potent antimicrobial producers from the primary screening among which 46.34% were effective against Gram-positive and 12.19% against Gram-negative test organisms. Isolate C7 (Micromonospora spp.) showed the best broad-spectrum antimicrobial activity during secondary screening. A total of 11 different types of pigments were observed to be produced by different isolates, of which, the yellow pigment was the most prominent. The association between elevation, pH, and pigment with the antimicrobial production was found to be insignificant. This finding can be of importance for further investigation towards obtaining broad-spectrum antibiotics for therapeutic purpose.
