Isolation and Characterization of Lytic Bacteriophage Against Common Pathogenic Bacteria

Asma S. Alilesh; Marwa E. Elwash; Tasneem M. Alswehly; Khawla A. Aween

doi:10.54172/21srcz92

Authors

Asma S. Alilesh Department of Genetics and Biotechnology, Faculty of Science, University of Misurata, Libya Author
Marwa E. Elwash Department of Genetics and Biotechnology, Faculty of Science, University of Misurata, Libya Author
Tasneem M. Alswehly Department of Genetics and Biotechnology, Faculty of Science, University of Misurata, Libya Author
Khawla A. Aween Department of Genetics and Biotechnology, Faculty of Science, University of Misurata, Libya Author

DOI:

https://doi.org/10.54172/21srcz92

Keywords:

Bacteriophages, Sewage water, Escherichia coli, Staphylococcus aureus, Antibacterial Agent

Abstract

Regarding their specificity, bacteriophages have been widely investigated to combat bacterial infections. Phage therapy is proposed as a promising alternative antibacterial agent. The present study was conducted to isolate and characterize bacteriophages against clinical bacterial isolates from sewage water. Recovery of phage was high from the processed sewage water (66%) against tested bacterial hosts. Plaque assay revealed four different plaques morphology against Escherichia coli with high lytic activity. In contrast, one small morphology plaques were appeared against Staphylococcus aureus with low lytic activity. The mean phage titer of phage isolates was 7.7x10⁹ and 2.9x10¹¹ plaque forming unit/ml for S. aureus and E. coli respectively. The isolated phages showed a narrow host range when tested against 19 different bacterial isolates. The electron microscopy revealed that EC1 Phage has the typical morphology of the family Podoviridae, order Caudovirales. The isolated bacteriophages need to be further characterized at the molecular level and tested in vivo to be used in one of the bacteriophage applications.

References

Al-Anany, A. M., Hooey, P. B., Cook, J. D., Burrows, L. L., Martyniuk, J., Hynes, A. P., & German, G. J. 2023. Phage Therapy in the Management of Urinary Tract Infections: A Comprehensive Systematic Review. PHAGE (New Rochelle, N.Y.), 4(3),112–127.

Al-Saffar, M. 2019. Isolation and characterization of lytic bacteriophages infecting Acinetobacter Baunannii from sewage water in babylon province, Iraq. Biochem. Cell. Arch., 19(1): 1881-1888.

Carey-Smith, G. V., Billington, C., Cornelius, A. J., Hudson, J. A., & Heinemann, J. A. 2006. Isolation and characterization of bacteriophages infecting Salmonella spp. FEMS microbiology letters, 258(2),182–186.

Chen, Y., Sun, E., Song, J., Yang, L., & Wu, B. 2018. Complete Genome Sequence of a Novel T7-Like Bacteriophage from a Pasteurella multocida Capsular Type A Isolate. Current microbiology, 75(5),574–579.

Duarte, J., Máximo, C., Costa, P., Oliveira, V., Gomes, N.C.M., Romalde, J.L., Pereira, C., & Almeida, A. 2024. Potential of an Isolated Bacteriophage to Inactivate Klebsiella pneumoniae: Preliminary Studies to Control Urinary Tract Infections. Antibiotics .13(2), 195.

Esmael, A., Azab, E., Gobouri, A. A., Nasr-Eldin, M. A., Moustafa, M. M. A., Mohamed, S. A., Badr, O. A. M., & Abdelatty, A. M. 2021. Isolation and Characterization of Two Lytic Bacteriophages Infecting a Multi-Drug Resistant Salmonella Typhimurium and Their Efficacy to Combat Salmonellosis in Ready-to-Use Foods. Microorganisms, 9(2),423.

Gallet, R., Kannoly, S., & Wang, I. N. 2011. Effects of bacteriophage traits on plaqueformation. BMC Microbiology, 11,181.

Kaur, S., Harjai, K., & Chhibber, S. 2012. Methicillin-resistant Staphylococcus aureus phage plaque size enhancement using sublethal concentrations of antibiotics. Applied and environmental microbiology, 78(23), 8227–8233.

Le, T., Nang, S. C., Zhao, J., Yu, H. H., Li, J., Gill, J. J., Liu, M., & Aslam, S. 2023. Therapeutic Potential of Intravenous Phage as Standalone Therapy for Recurrent Drug-Resistant Urinary Tract Infections. Antimicrobial agents and chemotherapy, 67(4), e0003723.

Masoud, S., Refat, H., Sayed, N., Abd-El Aal, M., Dosocky, A., Mohammed, Z., Abdel-Wahab, M., Mekawy, D., Ali, A., Atya, B., Welliam, K., Abd El-Baky, R. and Hashem, Z. 2020. Application of bacteriophages isolated from sewage to control urinary O157:H7 Escherichia coli and several bacterial uropathogens. Novel research in microbiology journal, 4(5): 1005-1014.

Mattila, S., Ruotsalainen, P., & Jalasvuori, M. 2015. On-Demand Isolation of Bacteriophages Against Drug-Resistant Bacteria for Personalized Phage Therapy. Frontiers in microbiology, 6,1271.

Megha, P.S., Murugan S. and Harikumar, P.S. 2017. Isolation and Characterization of Lytic Coliphages from Sewage Water. Journal of pure applied microbiology, 11(1): 559-565.

Othman, B. A., Askora, A., & Abo-Senna, A. S. 2015. Isolation and characterization of a Siphoviridae phage infecting Bacillus megaterium from a heavily trafficked holy site in Saudi Arabia. Folia Microbiologica, 60(4), 289–295.

Qamar, H., Owais, M., Chauhan, D. and Rehman, S.2019. Isolation of bacteriophages from untreated sewage water against multi-drug resistant E. coli - An initiative to fight against drug resistance .Research square,10(21203): 1-15

Rasool, M. H., Yousaf, R., Siddique, A. B., Saqalein, M., & Khurshid, M. 2016. Isolation, Characterization, and Antibacterial Activity of Bacteriophages Against Methicillin-Resistant Staphylococcus aureus in Pakistan. Jundishapur journal of microbiology, 9(10),e36135.

Ross, A., Ward, S., & Hyman, P. 2016. More Is Better: Selecting for Broad Host Range Bacteriophages. Frontiers in microbiology, 7,1352.

Sambrook, J., Fritsch, E. R., & Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual (2nd ed.). Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.

Shende, R. K., Hirpurkar, S. D., Sannat, C., Rawat, N., & Pandey, V. 2017. Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens. Veterinary world, 10(8), 973–978.

Sjahriani, T., Wasito, E. B., & Tyasningsih, W. 2021. Isolation and Identification of Escherichia coli O157:H7 Lytic Bacteriophage from Environment Sewage. International journal of food science, 2021,7383121.

Spanakis, E., & Horne, M. T. 1987. Co-adaptation of Escherichia coli and coliphage lambda vir in continuous culture. Journal of general microbiology, 133(2), 353–360.

Ullah, A., Qamash, T., Khan, F. A., Sultan, A., Ahmad, S., Abbas, M., Khattak, M. A. K., Begum, N., Din, S. U., Jamil, J., & Kalsoom 2021. Characterization of a Coliphage AS1 isolated from sewage effluent in Pakistan. Brazilian journal of biology - Revista brasleira de biologia, 82, e240943.

Vera-Mansilla, J., Sánchez, P., Silva-Valenzuela, C. A., & Molina-Quiroz, R. C. 2022. Isolation and Characterization of Novel Lytic Phages Infecting Multidrug-Resistant Escherichia coli. Microbiology spectrum, 10(1), e0167821.

Yu, Y. P., Gong, T., Jost, G., Liu, W. H., Ye, D. Z., & Luo, Z. H. 2013. Isolation and characterization of five lytic bacteriophages infecting a Vibrio strain closely related to Vibrio Owensii. FEMS microbiology letters, 348(2),112–119.