Isolation and Characterization of Lytic Pseudomonas fluorescens Bacteriophage  isolated from milk

Rinratree  Wongyoo; Sirinthorn  Sunthornthummas; Komwit  Surachat; Thassanant  Atithep; Achariya  Rangsiruji; Siriruk  Sarawaneeyaruk; Onanong Pringsulaka

doi:10.59796/jcst.V13N2.2023.885

Authors

Rinratree Wongyoo Department of Microbiology, Faculty of Science, Srinakharinwirot University, Bangkok, 10110, Thailand
Sirinthorn Sunthornthummas National Biobank of Thailand (NBT), National Science and Technology Development Agency, Pathum Thani, Thailand 12120
Komwit Surachat Department of Biomedical Sciences and Biomedical Engineering, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla, Thailand 90110 & Translational Medicine Research Center, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla, Thailand 90110
Thassanant Atithep Frontier Research Center (FRC) Vidyasirimedhi Institute of Science and Technology (VISTEC). Wangchan Valley, Rayong, Thailand 21210
Achariya Rangsiruji Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok, 10110, Thailand
Siriruk Sarawaneeyaruk Department of Microbiology, Faculty of Science, Srinakharinwirot University, Bangkok, 10110, Thailand
Onanong Pringsulaka Department of Microbiology, Faculty of Science, Srinakharinwirot University, Bangkok, 10110, Thailand

DOI:

https://doi.org/10.59796/jcst.V13N2.2023.885

Keywords:

P. fluorescens, bacteriophage, biocontrol

Abstract

Phage-based biocontrol is an alternative method for preventing and controlling the occurrence of Pseudomonas spp. in food products. However, the use of bacteriophages to control heat-stable protease Pseudomonas fluorescens is still rare. The objectives of this study were to isolate lytic bacteriophages of P. fluorescens, and to evaluate their efficacy in controlling P. fluorescens at the laboratory level. Among these strains, only one phage, ΦTIS1, was isolated by using P. fluorescens TISTR 1887 as the host. Further analysis using electron microscopy indicated that ΦTIS1 belongs to the Myoviridae family. This phage ΦTIS1 was relatively stable at pH and temperature ranges of 4.0 to 12.0 and 4, 30, 37, and 45°C, respectively, after 1.5 h incubation. The partial ΦTIS1 genome was a linear double-stranded with a total length of 87,646 bp and a G + C content of 54.71%. In vitro studies of the effect of bacteriophages against P. fluorescens TISTR 1887 with phage ΦTIS1 using various multiplicity of infection (MOI) values showed a significant decrease in bacterial numbers during 6-12 hours of incubation, followed by bacterial regrowth. However, the phage was still able to significantly reduce the bacterial numbers compared to the control without phage. These findings suggest that phage ΦTIS1 has the potential to be an effective method for controlling the prevalence of spoilage-causing P. fluorescens strains in dairy industries.

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