Characterization of Bacteriophages from Sewage Water and Their Lytic Activity Against Mycobacterium smegmatis ATCC 14468

Rina Hidayati Pratiwi, Ade La Yusup, Tri Handayani Kurniati

Abstract

Bacteriophage-based inhibition of Mycobacterium tuberculosis offers a potential alternative for managing tuberculosis. In vitro testing generally utilizes Mycobacterium smegmatis as a surrogate model. This study aimed to isolate and characterize bacteriophages from wastewater capable of lysing M. smegmatis ATCC 14468, and to determine the optimal lytic activity from the combination of bacteriophage type and Multiplicity of Infection (MOI) doses. This research employed a 3x3 factorial randomized block design incorporating bacteriophage type and the MOI levels (0.01, 0.1, and 1) as independent variations. The dependent variable, lytic activity (PhageScore), was analyzed using a two-way ANOVA with 95% significance and Duncan's Multiple Range Test (DMRT) for post-hoc analysis. Three types of bacteriophages, namely Kc, Cm, and Mt, were successfully isolated from wastewater. These three bacteriophages shared similar characteristics: they were specific to M. smegmatis ATCC 14468, unable to infect host cells during the non-logarithmic (lag) phase and were only stable at pH 7. The stability of bacteriophage Mt could reach a temperature of 45°C, while bacteriophages Kc and Cm were stable at 37°C. The highest lytic activity was exhibited by bacteriophage Cm at a Multiplicity of Infection (MOI) of 1, yielding a PhageScore of 0.6920.

 

Keywords: Lytic Activity; Mycobacteriophage; Multiplicity of Infection; PhageScore; Phage Therapy; Tuberculosis; Wastewater.

 

DOI https://doi.org/10.55463/issn.1674-2974.53.7.11


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