A Novel Dhillonvirus Phage against Escherichia coli Bearing a Unique Gene of Intergeneric Origin
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strains and Growth Conditions
2.2. Sample Collections
2.3. Phage Isolation and Enrichment
2.4. Bacteriophage Plaque Assay
2.5. Phage DNA Extraction
2.6. Genome Sequencing
2.7. Phage Genome Assembly and Characterization
2.8. Transmission Electron Microscopy (TEM) Methodology
2.9. One-Step Growth Curve and Adsorption Assay
2.10. Thermal and pH Stability
2.11. Host Range Determination
2.12. Phylogenetic Analysis
3. Results
3.1. Genome Characterization
3.2. Genome Organization
3.3. Bacteriophage Plaque Formation and Morphology
3.4. One-Step Growth Curve and Adsorption Assay
3.5. Thermal and PH Stability of the Phage
3.6. Host Range Determination
3.7. Phylogenetic Analysis
3.8. Phylogenetic Analysis of a Unique Putative Tail Fiber Protein
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample No. | Bacterial Strains/ Isolates * | Samples Source | Antibiotics Resistance $ | Phylogroup & | Phage Lytic Activity ¶ |
---|---|---|---|---|---|
1 | Escherichia coli 823 | Wastewater treatment plant | S | A | + |
2 | Escherichia coli 668 | Wastewater treatment plant | S | B1 | + |
3 | Escherichia coli 824 | Wastewater treatment plant | S | B2 | − |
4 | Escherichia coli 663 | Wastewater treatment plant | S | D | |
5 | Escherichia coli 792 | Wastewater treatment plant | MDR | A | − |
6 | Escherichia coli 494 | Wastewater treatment plant | MDR | B1 | − |
7 | Escherichia coli 810 | Wastewater treatment plant | MDR | B2 | − |
8 | Escherichia coli 638 | Wastewater treatment plant | MDR | D | − |
9 | Escherichia coli 640 | Wastewater treatment plant | R | A | − |
10 | Escherichia coli 643 | Wastewater treatment plant | R | B1 | − |
11 | Escherichia coli 809 | Wastewater treatment plant | R | B2 | − |
12 | Escherichia coli 635 | Wastewater treatment plant | R | D | − |
13 | Escherichia coli 865 | Hospital wastewater | WT | A | + |
14 | Escherichia coli 866 | Hospital wastewater | WT | B1 | − |
15 | Escherichia coli 843 | Hospital wastewater | WT | B2 | − |
16 | Escherichia coli 580 | Hospital wastewater | WT | D | − |
17 | Escherichia coli 426 | Hospital wastewater | MDR | D | + |
18 | Escherichia coli 858 | Hospital wastewater | MDR | A | − |
19 | Escherichia coli 546 | Hospital wastewater | MDR | B2 | − |
20 | Escherichia coli 576 | Hospital wastewater | R | B2 | − |
21 | Escherichia coli 545 | Hospital wastewater | R | D | + |
22 | Escherichia coli 674 | Hospital wastewater | R | A | − |
23 | Escherichia coli 759 | River water | WT | A | − |
24 | Escherichia coli 774 | River water | WT | B1 | − |
25 | Escherichia coli 624 | River water | WT | B2 | − |
26 | Escherichia coli 769 | River water | WT | D | − |
27 | Escherichia coli 472 | River water | MDR | A | − |
28 | Escherichia coli 607 | River water | MDR | B1 | − |
29 | Escherichia coli 737 | River water | MDR | B2 | − |
30 | Escherichia coli 408 | River water | MDR | D | − |
31 | Escherichia coli 614 | River water | R | A | − |
32 | Escherichia coli 372 | River water | R | D | − |
33 | Escherichia coli 743 | River water | R | B2 | − |
34 | Escherichia coli 784 | River water | R | B1 | − |
35 | Escherichia coli 117 | Clinical | MDR | A | + |
36 | Escherichia coli 60 | Clinical | MDR | B2 | + |
37 | Escherichia coli 203 | Clinical | MDR | D | + |
38 | Escherichia coli 325 | Clinical | R | A | + |
39 | Escherichia coli 5 | Clinical | R | A | − |
40 | Escherichia coli 264 | Clinical | R | B1 | − |
41 | Escherichia coli 294 | Clinical | R | B2 | − |
42 | Escherichia coli 378 | Clinical | R | B2 | + |
43 | Escherichia coli 313 | Clinical | R | D | − |
44 | Escherichia coli 324 | Clinical | S | A | + |
45 | Escherichia coli 368 | Clinical | S | D | − |
46 | Escherichia coli 387 | Clinical | S | B2 | − |
47 | Escherichia coli 301 | Clinical | S | B1 | − |
48 | Escherichia coli 25922 | Reference strain from Becton Dickinson, France S.A.S | S | − | + |
49 | Escherichia coli 35218 | Reference strain from Becton Dickinson, France S.A.S | S | − | − |
50 | Escherichia coli 13846 | Reference strain from Becton Dickinson, France S.A.S | S | − | − |
51 | Escherichia coli O157:H7 | Clinical | MDR | − | − |
52 | Klebsiella pneumoniae 13883 | Reference strain from Becton Dickinson, France S.A.S | S | − | − |
53 | Klebsiella pneumoniae 700603 | Reference strain from Becton Dickinson, France S.A.S | S | − | − |
54 | Pseudomonas aeruginosa 27853 | Reference strain from Becton Dickinson, France S.A.S | S | − | − |
55 | Yersinia enterocolitica 9610 | Reference strain from Becton Dickinson, France S.A.S | S | − | − |
56 | Acinetobacter baumannii 17978 | Reference strain from Becton Dickinson, France S.A.S | S | − | − |
57 | Acinetobacter baumannii 19668 | Reference strain from Becton Dickinson, France S.A.S | S | − | − |
Phage Name | BLASTp Coverage (%) | BLASTp Percent Identity (%) | Genus | Family/ Morphotype | NCBI Accession Number |
---|---|---|---|---|---|
vB_EcoS_SA32RD | 98 | 72.5 | Tunavirus | Drexlerviridae | UIU27553.1 |
PGN6866 | 92 | 74.22 | Kuravirus | podoviruses | QKL16987.1 |
vB_EcoP_YF01 | 92 | 72.89 | Kuravirus | podoviruses | WBF04932.1 |
IME267 | 92 | 72.44 | Kuravirus | podoviruses | YP_010673185.1 |
MLP3 | 92 | 72.89 | Kuravirus | podoviruses | UEN68517.1 |
vB_EcoP-101114UKE3 | 92 | 72.44 | Kuravirus | podoviruses | YP_010673043.1 |
νB_EcoP_SU7 | 92 | 72 | Kuravirus | podoviruses | YP_010672804.1 |
vB_EcoS_011D5 | 99 | 38.7 | Dhillonvirus | siphoviruses | QMP82830.1 |
vB_EcoS_L-h 1M | 99 | 37.79 | Dhillonvirus | siphoviruses | UNY42316.1 |
vB_EcoS_SA30RD | 72 | 48.02 | Tunavirus | Drexlerviridae | UIU27628.1 |
vB_EcoS_Chapo | 72 | 47.46 | Tunavirus | Drexlerviridae | QLF82390.1 |
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Vasileiadis, A.; Bozidis, P.; Konstantinidis, K.; Kesesidis, N.; Potamiti, L.; Kolliopoulou, A.; Beloukas, A.; Panayiotidis, M.I.; Havaki, S.; Gorgoulis, V.G.; et al. A Novel Dhillonvirus Phage against Escherichia coli Bearing a Unique Gene of Intergeneric Origin. Curr. Issues Mol. Biol. 2024, 46, 9312-9329. https://doi.org/10.3390/cimb46090551
Vasileiadis A, Bozidis P, Konstantinidis K, Kesesidis N, Potamiti L, Kolliopoulou A, Beloukas A, Panayiotidis MI, Havaki S, Gorgoulis VG, et al. A Novel Dhillonvirus Phage against Escherichia coli Bearing a Unique Gene of Intergeneric Origin. Current Issues in Molecular Biology. 2024; 46(9):9312-9329. https://doi.org/10.3390/cimb46090551
Chicago/Turabian StyleVasileiadis, Anastasios, Petros Bozidis, Konstantinos Konstantinidis, Nikolaos Kesesidis, Louiza Potamiti, Anna Kolliopoulou, Apostolos Beloukas, Mihalis I. Panayiotidis, Sophia Havaki, Vassilis G. Gorgoulis, and et al. 2024. "A Novel Dhillonvirus Phage against Escherichia coli Bearing a Unique Gene of Intergeneric Origin" Current Issues in Molecular Biology 46, no. 9: 9312-9329. https://doi.org/10.3390/cimb46090551