Isolation, characterization, and genomic analysis of three novel Herelleviridae family lytic bacteriophages against uropathogenic isolates of Staphylococcus saprophyticus

Abstract Background Staphylococcus saprophyticus (S. saprophyticus) is the second most prevalent etiological agent of urinary tract infections (UTIs) in young women. However, there is a paucity of data regarding its bacteriophage (phage). Therefore, this study was conducted to isolate and identify n...

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Main Authors: Hesamaddin Shirzad-Aski, Mahsa Yazdi, Alireza Mohebbi, Maryam Rafiee, Abbas Soleimani-Delfan, Alijan Tabarraei, Ezzat Allah Ghaemi
Format: Article
Language:English
Published: BMC 2025-03-01
Series:Virology Journal
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Online Access:https://doi.org/10.1186/s12985-025-02710-0
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author Hesamaddin Shirzad-Aski
Mahsa Yazdi
Alireza Mohebbi
Maryam Rafiee
Abbas Soleimani-Delfan
Alijan Tabarraei
Ezzat Allah Ghaemi
author_facet Hesamaddin Shirzad-Aski
Mahsa Yazdi
Alireza Mohebbi
Maryam Rafiee
Abbas Soleimani-Delfan
Alijan Tabarraei
Ezzat Allah Ghaemi
author_sort Hesamaddin Shirzad-Aski
collection DOAJ
description Abstract Background Staphylococcus saprophyticus (S. saprophyticus) is the second most prevalent etiological agent of urinary tract infections (UTIs) in young women. However, there is a paucity of data regarding its bacteriophage (phage). Therefore, this study was conducted to isolate and identify new lytic phages from municipal wastewater with the objective of increasing knowledge about phages and their genomes. Methods A total of 11 clinical isolates of S. saprophyticus and 30 wastewater samples were used to isolate three lytic phages (vB_SsapH-Golestan-100, vB_SsapH-Golestan101-M, and vB_SsapH-Golestan-105-M). The morphology, behavioral characteristics, and complete DNA genomes of these phages were analyzed. Results The microscopic images of the phages revealed that the sizes of their heads and tail lengths fell within the ranges of 90–111 nm and 234–266 nm, respectively. All phages exhibited high adsorption rates (99.5% in 15 min) and burst sizes (150–210 PFU per infected cell), with a potential for a narrow host range. Genomic analysis of Staphylococcus phages indicated a size of 136,433 base pairs (bp) with a guanine-cytosine (GC) content of 33.7% and 192 open reading frames (ORFs) for vB_SsapH-Golestan-100, 144,081 bp with a GC content of 29.6% and 205 ORFs for vB_SsapH-Golestan101-M, and 142,199 bp with a GC content of 30.6% and 203 ORFs for vB_SsapH-Golestan-105-M. A bioinformatics analysis indicated that all three phages belong to the Twortvirinae subfamily of Herelleviridae. Among the three phages, vB_SsapH-Golestan-100 exhibited the least similarity to previously known phages, with less than 21% similarity with its closest counterparts in genomic databases. Conclusions This study identified new phages that have the ability to destroy a broad range of S. saprophyticus isolates and may potentially be classified as a new genus and species within the Herelleviridae family in future studies.
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spelling doaj-art-6cd45a339b4a4e9e8e540fe5565694e22025-08-20T02:10:12ZengBMCVirology Journal1743-422X2025-03-0122111510.1186/s12985-025-02710-0Isolation, characterization, and genomic analysis of three novel Herelleviridae family lytic bacteriophages against uropathogenic isolates of Staphylococcus saprophyticusHesamaddin Shirzad-Aski0Mahsa Yazdi1Alireza Mohebbi2Maryam Rafiee3Abbas Soleimani-Delfan4Alijan Tabarraei5Ezzat Allah Ghaemi6Infectious Diseases Research Center, Golestan University of Medical SciencesLaboratory Sciences Research Center, Golestan University of Medical SciencesDepartment of Virology, School of Medicine, Iran University of Medical SciencesDepartment of Microbiology, School of Medicine, Golestan University of Medical SciencesDepartment of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of IsfahanInfectious Diseases Research Center, Golestan University of Medical SciencesLaboratory Sciences Research Center, Golestan University of Medical SciencesAbstract Background Staphylococcus saprophyticus (S. saprophyticus) is the second most prevalent etiological agent of urinary tract infections (UTIs) in young women. However, there is a paucity of data regarding its bacteriophage (phage). Therefore, this study was conducted to isolate and identify new lytic phages from municipal wastewater with the objective of increasing knowledge about phages and their genomes. Methods A total of 11 clinical isolates of S. saprophyticus and 30 wastewater samples were used to isolate three lytic phages (vB_SsapH-Golestan-100, vB_SsapH-Golestan101-M, and vB_SsapH-Golestan-105-M). The morphology, behavioral characteristics, and complete DNA genomes of these phages were analyzed. Results The microscopic images of the phages revealed that the sizes of their heads and tail lengths fell within the ranges of 90–111 nm and 234–266 nm, respectively. All phages exhibited high adsorption rates (99.5% in 15 min) and burst sizes (150–210 PFU per infected cell), with a potential for a narrow host range. Genomic analysis of Staphylococcus phages indicated a size of 136,433 base pairs (bp) with a guanine-cytosine (GC) content of 33.7% and 192 open reading frames (ORFs) for vB_SsapH-Golestan-100, 144,081 bp with a GC content of 29.6% and 205 ORFs for vB_SsapH-Golestan101-M, and 142,199 bp with a GC content of 30.6% and 203 ORFs for vB_SsapH-Golestan-105-M. A bioinformatics analysis indicated that all three phages belong to the Twortvirinae subfamily of Herelleviridae. Among the three phages, vB_SsapH-Golestan-100 exhibited the least similarity to previously known phages, with less than 21% similarity with its closest counterparts in genomic databases. Conclusions This study identified new phages that have the ability to destroy a broad range of S. saprophyticus isolates and may potentially be classified as a new genus and species within the Herelleviridae family in future studies.https://doi.org/10.1186/s12985-025-02710-0Staphylococcus saprophyticusLytic bacteriophageHerelleviridaeWhole genome sequencingUrinary tract infection
spellingShingle Hesamaddin Shirzad-Aski
Mahsa Yazdi
Alireza Mohebbi
Maryam Rafiee
Abbas Soleimani-Delfan
Alijan Tabarraei
Ezzat Allah Ghaemi
Isolation, characterization, and genomic analysis of three novel Herelleviridae family lytic bacteriophages against uropathogenic isolates of Staphylococcus saprophyticus
Virology Journal
Staphylococcus saprophyticus
Lytic bacteriophage
Herelleviridae
Whole genome sequencing
Urinary tract infection
title Isolation, characterization, and genomic analysis of three novel Herelleviridae family lytic bacteriophages against uropathogenic isolates of Staphylococcus saprophyticus
title_full Isolation, characterization, and genomic analysis of three novel Herelleviridae family lytic bacteriophages against uropathogenic isolates of Staphylococcus saprophyticus
title_fullStr Isolation, characterization, and genomic analysis of three novel Herelleviridae family lytic bacteriophages against uropathogenic isolates of Staphylococcus saprophyticus
title_full_unstemmed Isolation, characterization, and genomic analysis of three novel Herelleviridae family lytic bacteriophages against uropathogenic isolates of Staphylococcus saprophyticus
title_short Isolation, characterization, and genomic analysis of three novel Herelleviridae family lytic bacteriophages against uropathogenic isolates of Staphylococcus saprophyticus
title_sort isolation characterization and genomic analysis of three novel herelleviridae family lytic bacteriophages against uropathogenic isolates of staphylococcus saprophyticus
topic Staphylococcus saprophyticus
Lytic bacteriophage
Herelleviridae
Whole genome sequencing
Urinary tract infection
url https://doi.org/10.1186/s12985-025-02710-0
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