Isolation and Identification of Actinomycetes with Antibacterial Activity from Soil Samples Around Kandy, Sri Lanka
Abstract
Commercially exploited antibiotics are originally isolated from Actinomycetes. Even so, Actinomycetes are mostly underestimated for their antibacterial potential. This study aimed to isolate and identify Actinomycetes with antibacterial activity from soil samples around Kandy Sri Lanka. Soils were taken from five different habitats (waste disposal (WS), originally cultivated (OR), riverbanks (RB), pasture (PS), and rhizospheric (RH)). Serially diluted samples were grown on an Actinomycetes isolation agar medium and screened for antibacterial activity against Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC25923), Pseudomonas aeruginosa (ATCC27853) and Klebsiella pneumoniae (ATCC13883) using perpendicular streak method. Isolates with inhibitory activity were subjected to secondary screening. 19 actinomycetes isolates were identified of which 10 isolates showed inhibitory activity during preliminary screening. During the secondary screening, two isolates from RB site and one isolate from OR site expressed broad-spectrum antibacterial activity while the other five isolates from RB, OR, WS, and PS sites inhibited either Escherichia coli or Staphylococcus aureus. The soil samples are a promising source of novel antibacterial compounds and thus these findings can be used for further investigations in developing broad-spectrum antibiotics for therapeutic targets in the future.
Keywords:
Actinomycetes, soil, antibacterial activity, S. aureus, E. coli
References
Fiedler HP, Bruntner C, Bull AT, Ward AC, Goodfellow M, Potterat O, et al. Marine actinomycetes as a source of novel secondary metabolites. Antonie Van Leeuwenhoek. 2005; 87:37–42.
2. Das R, Romi W, Das R, Et al. Antimicrobial potentiality of actinobacteria isolated from two microbiologically unexplored forest ecosystems of Northeast India. BMC Microbiol 2018:18-71.
3. Rifaat HM. The biodiversity of actinomycetes in the river Nile exhibiting antifungal activity. J Medit err Ecol. 2003; 4:5–7.
4. Ubramani R, Aalbersberg W. Marine actinomycetes: an ongoing source of novel bioactive metabolites. Microbiol Res. 2012; 167:571–80.
5. Liu X, Ashforth E, Ren B, Song F, Dai H, Liu M,Wang J, Xie Q, Zhang L. Bioprospecting Microbial Natural Product Libraries from the Marine Environment for Drug Discovery. J Antibiot. 2010; 63: 415-422.
6. Farda B, Djebaili R, Vaccarelli I, Del Gallo M, Pellegrini M. Actinomycetes from Caves: An Overview of Their Diversity, Biotechnological Properties, and Insights for Their Use in Soil Environments. Microorganisms. 2022; 10(2): 453.
7. Kitouni M, Boudemagh A, Oulmi L, Reghioua S, Boughachiche F, Zerizer H, et al. Isolation of actinomycetes producing bioactive substances from water, soil and tree bark samples of the north-east of Algeria. J Mycol Med. 2005; 15:45–51.
Barka EA, Vatsa P, Sanchez L, Gaveau-Vaillant N, Jacquard C, Klenk H-P, et al. Taxonomy, physiology, and natural products of actinobacteria. Microbiol Mol Biol Rev. 2016; 80:1–43
9. Sharma P, Kalita MC, Thakur D. Broad spectrum antimicrobial activity of forest-derived soil actinomycete, Nocardia sp. PB-52. Front Microbiol. 2016; 7:1–17.
10. Lee LH, Zainal N, Azman AS, Eng SK, Goh BH, Yin WF, et al. Diversity and antimicrobial activities of actinobacteria isolated from tropical mangrove sediments in Malaysia. Sci World J. 2014; 2014:1–14
11. De Simeis D, Serra S. Actinomycetes: A Never-Ending Source of Bioactive Compounds-An Overview on Antibiotics Production. Antibiotics (Basel). 2021 Apr 22;10(5):483.
12. Das R, Romi W, Das R, Sharma HK, Thakur D. Antimicrobial Potentiality of Actinobacteria Isolated from Two Microbiologically Unexplored Forest Ecosystems of Northeast India. BMC Microbiol. 2018, 18-71.
13. Salaria N. Isolation and Screening of Actinomycetes from Soil Sample of Dal Lake (Kashmir) Against Selected Pathogens; 2018.
14. Lee LH, Zainal N, Azman AS, Eng SK, Goh BH, Yin WF, et al. Diversity and antimicrobial activities of actinobacteria isolated from tropical mangrove sediments in Malaysia. Sci World J.2014:1–14.
15. Tilahun R. Isolation of Antibiotics Producing Actinobacteria from Biowaste Soil Samples. Int. J. Microbiol. Biotechnol. 2020, 203-206.
16. Qin S, Li J, Chen HH, Zhao GZ, Zhu WY, Jiang CL, et al. Isolation, diversity, and antimicrobial activity of rare actinobacteria from medicinal plants of tropical rain forests in Xishuangbanna China. Appl Environ Microbiol. 2009; 75:6176–86.
17. Passari AK, Mishra VK, Saikia R, Gupta VK, Singh BP. Isolation, abundance and phylogenetic affiliation of endophytic actinomycetes associated with medicinal plants and screening for their in vitro antimicrobial biosynthetic potential. Front Microbiol. 2015; 6:1–13.
18. Nasrabadi R, Greiner R, Alikhani H, Hamedi J, Yakhchali B. Distribution of Actinomycetes in Different Soil Ecosystems and Effect of Media Composition on Extracellular Phosphatase Activity. J Soil Science and Plamt nutrition. 2013; 223-236.
19. Rana S, Salam MD. Antimicrobial potential of actinomycetes isolated from soil samples of Punjab, India. J Microbiol Exp. 2014;1(2):63-68.
20. Wahab A, and et al, Isolation and identification of actinomycetes isolated from karachi soil and screening of antimicrobial compounds, International Journal of Current Research. Https://www.journalcra.com/article/isolation-and-identification-actinomycetes-isolated-karachi-soil-and-screening-antimicrobial (accessed 2022-08-17).
21. Milshteyn A, Schneider JS, Brady SF. Mining the metabiome: identifying novel natural products from microbial communities. Chem Biol. 2015; 21:1211–23.
22. Gunasinghe YHKIS. and Edirisinghe EAAD. Soil Actinomycetes: potential source of novel antimicrobial substances. Proceedings of the 4th International Research Symposium on Pure and Applied Sciences, Faculty of Science, University of Kelaniya, Sri Lanka (2019) ;4-67.
23. Hughes D, Karlén A. Discovery and Preclinical Development of New Antibiotics. Ups. J. Med. Sci. 2014, 162-169.
24. Ayuso-Sacido A, Genilloud O. New PCR primers for the screening of NRPS and PKS-I systems in actinomycetes: detection and distribution of these biosynthetic gene sequences in major taxonomic groups. Microb Ecol. 2005; 49:10–24
25. Aryal S. Spread Plate Technique- Principle, Procedure and Uses. Microbiology Info.com, 2016.
26. Burke V, Barnes MW. The Cell Wall and The Gram Reaction. J Bacteriol. 1929; 18(2): 69-92.
27. Velho-Pereira S, Kamat NM. Antimicrobial Screening of Actinobacteria using a Modified Cross-Streak Method. Indian J Pharm Sci. 2011 Mar;73(2):223-8.
28. Luisa B.G., Actinomycetes and Biotechnology, 1st edition, (1988).
29. Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal. 2016 Apr;6(2):71-79.
30. Gottlieb A, Stein GY, Ruppin E, Sharan R. PREDICT: A Method for Inferring Novel Drug Indications with Application to Personalized Medicine. Mol. Syst. Biol. 2011, 7, 496.
31. Sapkota A, Thapa A, Budhathoki A, Sainju M, Shrestha P, Aryal S. Isolation, Characterization, and Screening of Antimicrobial-Producing Actinomycetes from Soil Samples. Int J Microbiol. 2020; 2020:2716584
32. Gottlieb A, Stein GY, Ruppin E, Sharan R. PREDICT: a method for inferring novel drug indications with application to personalized medicine. Mol Syst Biol. 2011; 7:496.
33. Singh LS, Sharma H & Talukdar NC. Production of potent antimicrobial agent by actinomycete, Streptomyces sannanensis strain SU118 isolated from phoomdi in Loktak Lake of Manipur, India. BMC Microbiol 14, (2014); 278.
2. Das R, Romi W, Das R, Et al. Antimicrobial potentiality of actinobacteria isolated from two microbiologically unexplored forest ecosystems of Northeast India. BMC Microbiol 2018:18-71.
3. Rifaat HM. The biodiversity of actinomycetes in the river Nile exhibiting antifungal activity. J Medit err Ecol. 2003; 4:5–7.
4. Ubramani R, Aalbersberg W. Marine actinomycetes: an ongoing source of novel bioactive metabolites. Microbiol Res. 2012; 167:571–80.
5. Liu X, Ashforth E, Ren B, Song F, Dai H, Liu M,Wang J, Xie Q, Zhang L. Bioprospecting Microbial Natural Product Libraries from the Marine Environment for Drug Discovery. J Antibiot. 2010; 63: 415-422.
6. Farda B, Djebaili R, Vaccarelli I, Del Gallo M, Pellegrini M. Actinomycetes from Caves: An Overview of Their Diversity, Biotechnological Properties, and Insights for Their Use in Soil Environments. Microorganisms. 2022; 10(2): 453.
7. Kitouni M, Boudemagh A, Oulmi L, Reghioua S, Boughachiche F, Zerizer H, et al. Isolation of actinomycetes producing bioactive substances from water, soil and tree bark samples of the north-east of Algeria. J Mycol Med. 2005; 15:45–51.
Barka EA, Vatsa P, Sanchez L, Gaveau-Vaillant N, Jacquard C, Klenk H-P, et al. Taxonomy, physiology, and natural products of actinobacteria. Microbiol Mol Biol Rev. 2016; 80:1–43
9. Sharma P, Kalita MC, Thakur D. Broad spectrum antimicrobial activity of forest-derived soil actinomycete, Nocardia sp. PB-52. Front Microbiol. 2016; 7:1–17.
10. Lee LH, Zainal N, Azman AS, Eng SK, Goh BH, Yin WF, et al. Diversity and antimicrobial activities of actinobacteria isolated from tropical mangrove sediments in Malaysia. Sci World J. 2014; 2014:1–14
11. De Simeis D, Serra S. Actinomycetes: A Never-Ending Source of Bioactive Compounds-An Overview on Antibiotics Production. Antibiotics (Basel). 2021 Apr 22;10(5):483.
12. Das R, Romi W, Das R, Sharma HK, Thakur D. Antimicrobial Potentiality of Actinobacteria Isolated from Two Microbiologically Unexplored Forest Ecosystems of Northeast India. BMC Microbiol. 2018, 18-71.
13. Salaria N. Isolation and Screening of Actinomycetes from Soil Sample of Dal Lake (Kashmir) Against Selected Pathogens; 2018.
14. Lee LH, Zainal N, Azman AS, Eng SK, Goh BH, Yin WF, et al. Diversity and antimicrobial activities of actinobacteria isolated from tropical mangrove sediments in Malaysia. Sci World J.2014:1–14.
15. Tilahun R. Isolation of Antibiotics Producing Actinobacteria from Biowaste Soil Samples. Int. J. Microbiol. Biotechnol. 2020, 203-206.
16. Qin S, Li J, Chen HH, Zhao GZ, Zhu WY, Jiang CL, et al. Isolation, diversity, and antimicrobial activity of rare actinobacteria from medicinal plants of tropical rain forests in Xishuangbanna China. Appl Environ Microbiol. 2009; 75:6176–86.
17. Passari AK, Mishra VK, Saikia R, Gupta VK, Singh BP. Isolation, abundance and phylogenetic affiliation of endophytic actinomycetes associated with medicinal plants and screening for their in vitro antimicrobial biosynthetic potential. Front Microbiol. 2015; 6:1–13.
18. Nasrabadi R, Greiner R, Alikhani H, Hamedi J, Yakhchali B. Distribution of Actinomycetes in Different Soil Ecosystems and Effect of Media Composition on Extracellular Phosphatase Activity. J Soil Science and Plamt nutrition. 2013; 223-236.
19. Rana S, Salam MD. Antimicrobial potential of actinomycetes isolated from soil samples of Punjab, India. J Microbiol Exp. 2014;1(2):63-68.
20. Wahab A, and et al, Isolation and identification of actinomycetes isolated from karachi soil and screening of antimicrobial compounds, International Journal of Current Research. Https://www.journalcra.com/article/isolation-and-identification-actinomycetes-isolated-karachi-soil-and-screening-antimicrobial (accessed 2022-08-17).
21. Milshteyn A, Schneider JS, Brady SF. Mining the metabiome: identifying novel natural products from microbial communities. Chem Biol. 2015; 21:1211–23.
22. Gunasinghe YHKIS. and Edirisinghe EAAD. Soil Actinomycetes: potential source of novel antimicrobial substances. Proceedings of the 4th International Research Symposium on Pure and Applied Sciences, Faculty of Science, University of Kelaniya, Sri Lanka (2019) ;4-67.
23. Hughes D, Karlén A. Discovery and Preclinical Development of New Antibiotics. Ups. J. Med. Sci. 2014, 162-169.
24. Ayuso-Sacido A, Genilloud O. New PCR primers for the screening of NRPS and PKS-I systems in actinomycetes: detection and distribution of these biosynthetic gene sequences in major taxonomic groups. Microb Ecol. 2005; 49:10–24
25. Aryal S. Spread Plate Technique- Principle, Procedure and Uses. Microbiology Info.com, 2016.
26. Burke V, Barnes MW. The Cell Wall and The Gram Reaction. J Bacteriol. 1929; 18(2): 69-92.
27. Velho-Pereira S, Kamat NM. Antimicrobial Screening of Actinobacteria using a Modified Cross-Streak Method. Indian J Pharm Sci. 2011 Mar;73(2):223-8.
28. Luisa B.G., Actinomycetes and Biotechnology, 1st edition, (1988).
29. Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal. 2016 Apr;6(2):71-79.
30. Gottlieb A, Stein GY, Ruppin E, Sharan R. PREDICT: A Method for Inferring Novel Drug Indications with Application to Personalized Medicine. Mol. Syst. Biol. 2011, 7, 496.
31. Sapkota A, Thapa A, Budhathoki A, Sainju M, Shrestha P, Aryal S. Isolation, Characterization, and Screening of Antimicrobial-Producing Actinomycetes from Soil Samples. Int J Microbiol. 2020; 2020:2716584
32. Gottlieb A, Stein GY, Ruppin E, Sharan R. PREDICT: a method for inferring novel drug indications with application to personalized medicine. Mol Syst Biol. 2011; 7:496.
33. Singh LS, Sharma H & Talukdar NC. Production of potent antimicrobial agent by actinomycete, Streptomyces sannanensis strain SU118 isolated from phoomdi in Loktak Lake of Manipur, India. BMC Microbiol 14, (2014); 278.
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How to Cite
Navoda Bandara, and Tennakoon T.M.I.U.K. “Isolation and Identification of Actinomycetes With Antibacterial Activity from Soil Samples Around Kandy, Sri Lanka”. Current Research in Pharmaceutical Sciences, Vol. 13, no. 3, Dec. 2023, pp. 118-25, doi:10.24092/CRPS.2023.130301.
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Research Articles
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
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