Identification and Characterization of a Bacteriophage with Lytic Activity against Multidrug Resistant E. coli
Keywords:Pathogenic E. coli, bacteriophages, phage therapy, sewage
Escherichia coli strains are increasingly becoming resistant to antibiotics and emerging globally. Bacteriophage is re-explored for the development of an alternative and safe agent to control a bacterial infection, especially with multi-drug resistant (MDR) bacteria. Here, we reported the identification of AAA1 phage from a sewage site near Baghdad’s Medical City. Morphological analysis using Transmission Electron Microscopy (TEM) suggested that the AAA1 phage had an icosahedral head of width ~ 70 nm and a non-contractile tail of ~ 400 nm belonging to the Siphoviridae family from the Caudovirales order. The plaque observation showed that plaques size and shape differ after prolonged incubation. The optimal multiplicity of infection was 0.1. The one-step growth curve was assessed and the result showed that AAA1 latent period was 10 minutes; burst size was 93PFU/cell and the burst period of 30 minutes. Based on characteristics, AAA1 phage is a potential candidate to control MDR E. coli infection.
J. Jang, H.-G. Hur, M. J. Sadowsky, M. N. Byappanahalli, T. Yan, and S. Ishii, "Environmental Escherichia coli : ecology and public health implications-a review," J. Appl. Microbiol., vol. 123, no. 3, pp. 570-581, Sep. 2017.
V. M. Vaughn, A. L. Hersh, and E. S. Spivak, "Antibiotic Overuse and Stewardship at Hospital Discharge: The Reducing Overuse of Antibiotics at Discharge Home Framework," Clin Infect Dis, vol. 3;74(9):1696-702, May 2022.
A. Ousaid, J. Akrim, and Y. Khayati, "Overuse of antibiotics as a key driver to antibiotic resistance in Morocco: A short review with potential solutions," Int Arab J Antimicrob Agents Internet, 2020,
A. H. Pishtiwan and K. M. Khadija, "Prevalence of blaTEM, blaSHV, and blaCTX-M genes among ESBL-producing Klebsiella pneumoniae and Escherichia coli isolated from thalassemia patients in Erbil, Iraq," Mediterr J Hematol Infect Dis, vol. 11, 2019.
D. R. Al-Guranie and S. M. Al-Mayahie, "Prevalence of E. coli ST131 among Uropathogenic E. coli isolates from Iraqi patients in Wasit Province, Iraq," Int J Microbiol, 2020.
H. H. Al-Hasnawy, M. R. Judi, and H. J. Hamza, "The dissemination of multidrug resistance (MDR) and extensively drug resistant (XDR) among uropathogenic E. coli (UPEC) isolates from urinary tract infection patients in babylon province, Iraq," Baghdad Sci J, vol. 16, pp. 986-992, 2019.
L. Poirel et al., "Antimicrobial resistance in Escherichia coli," Microbiol Spectr, vol. 6, pp. 6-4, 2018.
S. Saputra et al., "Antimicrobial resistance in clinical Escherichia coli isolated from companion animals in Australia," Vet Microbiol, vol. 211, pp. 43-50, 2017.
B. Fathima and A. C. Archer, "Bacteriophage therapy: recent developments and applications of a renaissant weapon," Res Microbiol, vol. 172, no. 6, 2021.
P. Wintachai and S. P. Voravuthikunchai, "Characterization of Novel Lytic Myoviridae Phage Infecting Multidrug-Resistant Acinetobacter baumannii and Synergistic Antimicrobial Efficacy between Phage and Sacha Inchi Oil," Pharmaceuticals, vol. 15, no. 3, 2022.
K. Hon, S. Liu, S. Camens, G. S. Bouras, A. J. Psaltis, and P.-J. Wormald, "APTC-EC-2A: A Lytic Phage Targeting Multidrug Resistant E. coli Planktonic Cells and Biofilms," Microorganisms, vol. 10, no. 1, 2022.
J. Vera-Mansilla, P. Sánchez, C. A. Silva-Valenzuela, and R. C. Molina-Quiroz, "Isolation and Characterization of Novel Lytic Phages Infecting Multidrug-Resistant Escherichia coli," Microbiol Spectr, vol. 10, no. 1, 2022.
K. Namdee, M. Khongkow, S. Boonrungsiman, N. Nittayasut, P. Asavarut, and S. Temisak, "Thermoresponsive Bacteriophage Nanocarrier as a Gene Delivery Vector Targeted to the Gastrointestinal Tract. Mol Ther - Nucleic Acids," vol. 12. pp. 33-44, Sep. 2018.
Z. D. Moye, J. Woolston, and A. Sulakvelidze, "Bacteriophage Applications for Food Production and Processing," Viruses, vol. Apr;10(4):205, 2018.
S. Royer, A. P. Morais, and D. W. Fonseca Batistão, "Phage therapy as strategy to face post-antibiotic era: a guide to beginners and experts," Arch Microbiol, vol. May; 203(4):1271-9, 2021.
C. Sieiro, L. Areal-Hermida, Á. Pichardo-Gallardo, R. Almuiña-González, T. Miguel, and S. Sánchez, "A hundred years of bacteriophages: Can phages replace antibiotics in agriculture and aquaculture?," Antibiotics, vol. 9, no. 8, 2020.
S. Aslam and R. T. Schooley, "What's Old Is New Again: Bacteriophage Therapy in the 21st Century," Antimicrob Agents Chemother, Dec. 2019.
V. Fanaei, M. Validi, B. Zamanzad, and A. Karimi, "Isolation and identification of specific bacteriophages against methicillin-resistant Staphylococcus aureus, extended-spectrum beta-lactamases-producing Escherichia coli, extended-spectrum beta-lactamases-producing Klebsiella pneumoniae, and multidrug-resistant Acinetobacter baumanniiin vitro," FEMS Microbiol Lett, vol. 1;368(19):fnab139, Oct. 2021.
A. A. Abdulhussein and B. O. Abdulsattar, "Isolation and characterization of two novel phages as a possible therapeutic alternative against multi-drug resistant E. coli," Gene Rep., vol. 28, p. 101644, Sep. 2022.
B. O. Abdulsattar, N. A. Al-Saryi, M. H. Abbas, and A.-R. A. Abdulhussein, "Isolation and purification of Escherichia coli bacteriophage from Tigris river, Baghdad, Iraq," Gene Rep, vol. 19, p. 100591, 2020.
C. Wei, J. Liu, A. N. Maina, F. B. Mwaura, J. Yu, and C. Yan, "Developing a bacteriophage cocktail for biocontrol of potato bacterial wilt," Virol Sin, vol. 32, no. 6, pp. 476-84, 2017.
P. Fu, Q. Zhao, L. Shi, Q. Xiong, Z. Ren, and H. Xu, "Identification and characterization of two bacteriophages with lytic activity against multidrug-resistant Escherichia coli," Virus Res, vol. 291, no. 198196, 2021.
R. Urwin and M. C. Maiden, "Multi-locus sequence typing: a tool for global epidemiology," Trends Microbiol, vol. 11, pp. 479-487, 2003.
W. G. Weisburg, S. M. Barns, D. A. Pelletier, and D. J. Lane, "16S ribosomal DNA amplification for phylogenetic study," J Bacteriol, vol. 173, pp. 697-703, 1991.
M. I. El-Amir, M. A. El-Feky, D. A. A. Elwafa, and E. A. Abd-Elmawgood, "Rapid diagnosis of neonatal sepsis by PCR for detection of 16S rRNA gene, while blood culture and PCR results were similar in E. coli-predominant EOS cases," Infect Drug Resist, vol. 12, p. 2703, 2019.
M. Allami, M. Bahreini, and M. R. Sharifmoghadam, "Antibiotic resistance, phylogenetic typing, and virulence genes profile analysis of uropathogenic Escherichia coli isolated from patients in southern Iraq," J Appl Genet, vol. 63, no. 2, pp. 401-12, 2022.
G. A.-M. SM, A.-G. DRT, H. AA, and B. ZA, "Prevalence of common carbapenemase genes and multidrug resistance among uropathogenic Escherichia coli phylogroup B2 isolates from outpatients in Wasit Province/Iraq," Plos One, vol. 17, no. 1, 2022.
R. N. Hasan, S. A. Jasim, and Y. H. Ali, "Detection of fimH, kpsMTII, hlyA, and traT genes in Escherichia coli isolated from Iraqi patients with cystitis," Gene Rep, vol. 26, no. 101468, 2022.
R. K. Mohammed and A. A. Ibrahim, "Distribution of dfrA1 and cat1 antibiotic resistance genes in uropathogenic Escherichia coli isolated from teens pregnant women in Iraq," Iraqi J Sci, vol. 3340-53, 2022.
H.-W. Ackermann, "5500 Phages examined in the electron microscope," Arch Virol, vol. 152, pp. 227-243, 2007.
F. Abdelrahman, N. Rezk, M. S. Fayez, M. Abdelmoteleb, R. Atteya, and M. Elhadidy, "Isolation, Characterization, and Genomic Analysis of Three Novel E. coli Bacteriophages That Effectively Infect E. coli O18," Microorganisms, vol. 10, no. 3, 2022.
E. W. Bumunang, T. A. McAllister, K. Stanford, H. Anany, Y. D. Niu, and C. N. Ateba, "Characterization of Non-O157 STEC infecting bacteriophages isolated from cattle faeces in North-West South Africa," Microorganisms, vol. 7, no. 12, 2019.
N. Chaudhary, C. Narayan, B. Mohan, and N. Taneja, "Characterization and in vitro activity of a lytic phage RDN37 isolated from community sewage water active against MDR Uropathogenic E. coli," Indian J Med Microbiol, vol. 39, no. 3, pp. 343-8, 2021.
M. M. Esmat, A. G. Abdelhamid, S. A. Abo-ELmaaty, M. A. Nasr-Eldin, M. G. Hassan, and A. A. Khattab, "Antibiotics and phage sensitivity as interventions for controlling Escherichia coli isolated from clinical specimens," J Pure Appl Microbiol, vol. 11, no. 4, pp. 1749-55, 2017.
Y.-K. Li, H. Chen, M. Shu, C. Zhong, Y. Bi, and H.-H. Yang, "Isolation, characterization and application of an alkaline resistant virulent bacteriophage JN01 against Escherichia coli O157: H7 in milk and beef," LWT, vol. 144, no. 111266, 2021.
M. Raj JR, V. R, H. P, B. U, and K. I, "T4-like Escherichia coli phages from the environment carry blaCTX-M," Lett Appl Microbiol, vol. 67, no. 1, pp. 9-14, 2018.
M. A. Nasr-Eldin, S. A. EL-Maaty, K. A. EL-Dougdoug, M. M. Hazaa, and A. H. Abdel-mageed, "Characterization and development of a phage cocktail for Escherichia coli causing gastrointestinal diseases," J Bas Env Sci, vol. 5, pp. 115-22, 2011.
A. Vieira, Y. J. Silva, A. Cunha, N. C. M. Gomes, H.-W. Ackermann, and A. Almeida, "Phage therapy to control multidrug-resistant Pseudomonas aeruginosa skin infections: in vitro and ex vivo experiments," Eur J Clin Microbiol Infect Dis, vol. 31, pp. 3241-3249, 2012.
C. Kocharunchitt, T. Ross, and D. L. McNeil, "Use of bacteriophages as biocontrol agents to control Salmonella associated with seed sprouts," Int J Food Microbiol, vol. 128, pp. 453-459, 2009.
J. Majewska et al., "Oral application of T4 phage induces weak antibody production in the gut and in the blood," Viruses, vol. 7, pp. 4783-4799, 2015.
S. T. Abedon, "Phage therapy dosing: The problem (s) with multiplicity of infection (MOI," Bacteriophage, vol. 6, p. 1220348, 2016.
Q. Peng and Y. Yuan, "Characterization of a newly isolated phage infecting pathogenic Escherichia coli and analysis of its mosaic structural genes," Sci Rep, vol. 8, pp. 1-10, 2018.
G.-H. Kim, J.-W. Kim, J. Kim, J. P. Chae, J.-S. Lee, and S.-S. Yoon, "Genetic analysis and characterization of a bacteriophage ØCJ19 active against enterotoxigenic Escherichia coli," Food Sci Anim Resour, vol. 40, p. 746, 2020.
S. T. Abedon, P. Hyman, and C. Thomas, "Experimental examination of bacteriophage latent-period evolution as a response to bacterial availability," Appl Env. Microbiol, vol. 69, no. 12, pp. 7499-506, 2003.
S. T. Abedon, T. D. Herschler, and D. Stopar, "Bacteriophage latent-period evolution as a response to resource availability," Appl Env. Microbiol, vol. 67, no. 9, pp. 4233-41, 2001.
A. S. Nilsson, "Phage therapy-constraints and possibilities," J Med Sci, vol. 119, no. 2, pp. 192-8, 2014.
A. J. Browne, M. G. Chipeta, G. Haines-Woodhouse, E. P. Kumaran, B. H. K. Hamadani, and S. Zaraa, "Global antibiotic consumption and usage in humans, 2000-18: a spatial modelling study," Lancet Planet Health, vol. 5, no. 12, 2021.
E. Y. Klein et al., "Global increase and geographic convergence in antibiotic consumption between 2000 and 2015," Proc Natl Acad Sci, vol. 115, pp. 3463-3470, 2018.
M. Alzaidi JR and A.S., "First Record of Dissemination of BLBLI-Resistant from Public Hospitals in Baghdad, Iraq," Open Microbiol J, vol. 16, no. 1, 2022.
M. H. G. Kanaan and H. T. Khashan, "Molecular typing, virulence traits and risk factors of pandrug-resistant Acinetobacter baumannii spread in intensive care unit centers of Baghdad city, Iraq," Rev Med Microbiol, vol. 33, no. 1, pp. 51-5, 2022.
H. A. Salman, S. M. S. Alsallameh, G. Muhamad, and Z. Taha, "Prevalence of Multi-Antibiotic Resistant Bacteria Isolated from Children with Urinary Tract Infection from Baghdad, Iraq," Microbiol Biotechnol Lett, vol. 50, no. 1, pp. 147-56, 2022.
F. M. Abbas, "Genetic characterization of OXA-1 β-lactamase producing carbapenem-resistant Klebsiella pneumoniae from Al-Hillah river water, Babylon Province, Iraq," J Genet Env. Resour Conserv, vol. 9, no. 1, pp. 197-201, 2021.
R. Barrangou, C. Fremaux, H. Deveau, M. Richards, P. Boyaval, and S. Moineau, "CRISPR Provides Acquired Resistance Against Viruses in Prokaryotes," Science, Mar. 2007.
T. Goldfarb, H. Sberro, E. Weinstock, O. Cohen, S. Doron, and Y. Charpak‐Amikam, "BREX is a novel phage resistance system widespread in microbial genomes," EMBO J, Jan. 2015.
P. Hyman and S. T. Abedon, "Bacteriophage Host Range and Bacterial Resistance," in Advances in Applied Microbiology, Elsevier, 2010, pp. 217-48.
S. J. Labrie, J. E. Samson, and S. Moineau, "Bacteriophage resistance mechanisms," Nat Rev Microbiol, May 2010.
C. Richter, J. T. Chang, and P. C. Fineran, "Function and Regulation of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) / CRISPR Associated (Cas) Systems," Viruses, Oct. 2012.
I. Yosef, M. Manor, R. Kiro, and U. Qimron, "Temperate and lytic bacteriophages programmed to sensitize and kill antibiotic-resistant bacteria," Proc Natl Acad Sci, Jun. 2015.
How to Cite
Copyright (c) 2023 Al-Mustansiriyah Journal of Science
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Articles accepted for publication in Al-Mustansiriyah Journal of Science (MJS) are protected under the Creative Commons Attribution 4.0 International License (CC BY-NC). Authors of accepted articles are requested to sign a copyright release form prior to their article being published. All authors must agree to the submission, sign copyright release forms, and agree to be included in any correspondence between MJS and the authors before submitting a work to MJS. For personal or educational use, permission is given without charge to print or create digital copies of all or portions of a MJS article. However, copies must not be produced or distributed for monetary gain. It is necessary to respect the copyright of any parts of this work that are not owned by MJS.