PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods

Carlos E Martinez-Soto; Stevan Cucić; Janet T Lin; Sarah Kirst; El Sayed Mahmoud; Cezar M Khursigara; Hany Anany

doi:10.3390/v13112120

PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods

Viruses. 2021 Oct 21;13(11):2120. doi: 10.3390/v13112120.

Authors

Carlos E Martinez-Soto^{1

2}, Stevan Cucić^{1

2}, Janet T Lin¹, Sarah Kirst¹, El Sayed Mahmoud³, Cezar M Khursigara¹, Hany Anany^{1

2}

Affiliations

¹ Guelph Research and Development Centre, Agriculture and Agri-Food Canada, Guelph, ON N1G 5C9, Canada.
² Department of Molecular and Cellular Biology, College of Biological Science, University of Guelph, Guelph, ON N1G 2W1, Canada.
³ Faculty of Applied Science and Technology, The Sheridan College Institute of Technology and Advanced Learning, Oakville, ON L6H 2L1, Canada.

Abstract

Bacteriophages are viruses that infect bacteria and are present in niches where bacteria thrive. In recent years, the suggested application areas of lytic bacteriophage have been expanded to include therapy, biocontrol, detection, sanitation, and remediation. However, phage application is constrained by the phage's host range-the range of bacterial hosts sensitive to the phage and the degree of infection. Even though phage isolation and enrichment techniques are straightforward protocols, the correlation between the enrichment technique and host range profile has not been evaluated. Agar-based methods such as spotting assay and efficiency of plaquing (EOP) are the most used methods to determine the phage host range. These methods, aside from being labor intensive, can lead to subjective and incomplete results as they rely on qualitative observations of the lysis/plaques, do not reflect the lytic activity in liquid culture, and can overestimate the host range. In this study, phages against three bacterial genera were isolated using three different enrichment methods. Host range profiles of the isolated phages were quantitatively determined using a high throughput turbidimetric protocol and the data were analyzed with an accessible analytic tool "PHIDA". Using this tool, the host ranges of 9 Listeria, 14 Salmonella, and 20 Pseudomonas phages isolated with different enrichment methods were quantitatively compared. A high variability in the host range index (HR_i) ranging from 0.86-0.63, 0.07-0.24, and 0.00-0.67 for Listeria, Salmonella, and Pseudomonas phages, respectively, was observed. Overall, no direct correlation was found between the phage host range breadth and the enrichment method in any of the three target bacterial genera. The high throughput method and analytics tool developed in this study can be easily adapted to any phage study and can provide a consensus for phage host range determination.

Keywords: bacteriophage; host range; isolation protocol; lytic phages; phage biocontrol; phage enrichment; phage therapy.

Publication types

Comparative Study
Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Bacteriophages / isolation & purification*
Bacteriophages / physiology*
Data Science / methods*
High-Throughput Screening Assays / methods*
Host Specificity*
Listeria / virology*
Pseudomonas / virology*
Salmonella / virology*
Software