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

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 constraine...

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Autores principales:	Carlos E. Martinez-Soto, Stevan Cucić, Janet T. Lin, Sarah Kirst, El Sayed Mahmoud, Cezar M. Khursigara, Hany Anany
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	bacteriophage isolation protocol host range phage biocontrol phage therapy phage enrichment Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/69bbd70a67d04b8f88bd16f329461b5e
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spelling	oai:doaj.org-article:69bbd70a67d04b8f88bd16f329461b5e2021-11-25T19:12:24ZPHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods10.3390/v131121201999-4915https://doaj.org/article/69bbd70a67d04b8f88bd16f329461b5e2021-10-01T00:00:00Zhttps://www.mdpi.com/1999-4915/13/11/2120https://doaj.org/toc/1999-4915Bacteriophages 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 <i>Listeria,</i> 14 <i>Salmonella</i>, and 20 <i>Pseudomonas</i> phages isolated with different enrichment methods were quantitatively compared. A high variability in the host range index (<i>HR<sub>i</sub>)</i> ranging from 0.86–0.63, 0.07–0.24, and 0.00–0.67 for <i>Listeria, Salmonella,</i> and <i>Pseudomonas</i> 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.Carlos E. Martinez-SotoStevan CucićJanet T. LinSarah KirstEl Sayed MahmoudCezar M. KhursigaraHany AnanyMDPI AGarticlebacteriophageisolation protocolhost rangephage biocontrolphage therapyphage enrichmentMicrobiologyQR1-502ENViruses, Vol 13, Iss 2120, p 2120 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	bacteriophage isolation protocol host range phage biocontrol phage therapy phage enrichment Microbiology QR1-502
spellingShingle	bacteriophage isolation protocol host range phage biocontrol phage therapy phage enrichment Microbiology QR1-502 Carlos E. Martinez-Soto Stevan Cucić Janet T. Lin Sarah Kirst El Sayed Mahmoud Cezar M. Khursigara Hany Anany PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods
description	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 <i>Listeria,</i> 14 <i>Salmonella</i>, and 20 <i>Pseudomonas</i> phages isolated with different enrichment methods were quantitatively compared. A high variability in the host range index (<i>HR<sub>i</sub>)</i> ranging from 0.86–0.63, 0.07–0.24, and 0.00–0.67 for <i>Listeria, Salmonella,</i> and <i>Pseudomonas</i> 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.
format	article
author	Carlos E. Martinez-Soto Stevan Cucić Janet T. Lin Sarah Kirst El Sayed Mahmoud Cezar M. Khursigara Hany Anany
author_facet	Carlos E. Martinez-Soto Stevan Cucić Janet T. Lin Sarah Kirst El Sayed Mahmoud Cezar M. Khursigara Hany Anany
author_sort	Carlos E. Martinez-Soto
title	PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods
title_short	PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods
title_full	PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods
title_fullStr	PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods
title_full_unstemmed	PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods
title_sort	phida: a high throughput turbidimetric data analytic tool to compare host range profiles of bacteriophages isolated using different enrichment methods
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/69bbd70a67d04b8f88bd16f329461b5e
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PHIDA: A High Throughput Turbidimetric Data Analytic Tool to Compare Host Range Profiles of Bacteriophages Isolated Using Different Enrichment Methods

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