Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant <i>Enterococcus faecalis</i>, the Causative Agent of Dental Abscess

<i>Background and Objectives</i>: <i>Enterococcus faecalis (E. faecalis)</i> is a primary pathogen responsible for dental abscesses, which cause inflammation and pain when trapped between the crown and soft tissues of an erupted tooth. Therefore, this study aims to use specif...

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Main Authors: Asmaa Ramadan, Mohamed O. Abdel-Monem, Noha K. El-Dougdoug, Alsayed E. Mekky, Shymaa A. Elaskary, Abdulaziz A. Al-Askar, Shimaa A Metwally, Ahmed F. El-Sayed, Gehad AbdElgayed, Ebrahim Saied, Mohamed Khedr
Format: Article
Language:English
Published: MDPI AG 2024-03-01
Series:Medicina
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Online Access:https://www.mdpi.com/1648-9144/60/3/501
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author Asmaa Ramadan
Mohamed O. Abdel-Monem
Noha K. El-Dougdoug
Alsayed E. Mekky
Shymaa A. Elaskary
Abdulaziz A. Al-Askar
Shimaa A Metwally
Ahmed F. El-Sayed
Gehad AbdElgayed
Ebrahim Saied
Mohamed Khedr
author_facet Asmaa Ramadan
Mohamed O. Abdel-Monem
Noha K. El-Dougdoug
Alsayed E. Mekky
Shymaa A. Elaskary
Abdulaziz A. Al-Askar
Shimaa A Metwally
Ahmed F. El-Sayed
Gehad AbdElgayed
Ebrahim Saied
Mohamed Khedr
author_sort Asmaa Ramadan
collection DOAJ
description <i>Background and Objectives</i>: <i>Enterococcus faecalis (E. faecalis)</i> is a primary pathogen responsible for dental abscesses, which cause inflammation and pain when trapped between the crown and soft tissues of an erupted tooth. Therefore, this study aims to use specific phages as an alternative method instead of classical treatments based on antibiotics to destroy multidrug-resistant <i>E. faecalis</i> bacteria for treating dental issues. <i>Materials and Methods</i>: In the current study, twenty-five bacterial isolates were obtained from infected dental specimens; only five had the ability to grow on bile esculin agar, and among these five, only two were described to be extensive multidrug-resistant isolates. <i>Results</i>: Two bacterial isolates, <i>Enterococcus faecalis</i> A.R.A.01 [ON797462.1] and <i>Enterococcus faecalis</i> A.R.A.02, were identified biochemically and through 16S rDNA, which were used as hosts for isolating specific phages. Two isolated phages were characterized through TEM imaging, which indicated that <i>E. faecalis</i>_phage-01 had a long and flexible tail, belonging to the family <i>Siphoviridae</i>, while <i>E</i>. <i>faecalis</i>_phage-02 had a contractile tail, belonging to the family <i>Myoviridae.</i> Genetically, two phages were identified through the PCR amplification and sequencing of the RNA ligase of <i>Enterococcus</i> phage vB_EfaS_HEf13, through which our phages shared 97.2% similarity with <i>Enterococcus</i> phage vB-EfaS-HEf13 based on BLAST analysis. Furthermore, through in silico analysis and annotations of the two phages’ genomes, it was determined that a total of 69 open reading frames (ORFs) were found to be involved in various functions related to integration excision, replication recombination, repair, stability, and defense. In phage optimization, the two isolated phages exhibited a high specific host range with <i>Enterococcus faecalis</i> among six different bacterial hosts, where <i>E. faecalis</i>_phage-01 had a latent period of 30 min with 115.76 PFU/mL, while <i>E</i>. <i>faecalis</i>_phage-02 had a latent period of 25 min with 80.6 PFU/mL. They were also characterized with stability at wide ranges of pH (4–11) and temperature (10–60 °C), with a low cytotoxic effect on the oral epithelial cell line at different concentrations (1000–31.25 PFU/mL). <i>Conclusions</i>: The findings highlight the promise of phage therapy in dental medicine, offering a novel approach to combating antibiotic resistance and enhancing patient outcomes. Further research and clinical trials will be essential to fully understand the therapeutic potential and safety profile of these bacteriophages in human populations.
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spelling doaj.art-a9ff62faf871443782d8eed6079f11262024-03-27T13:53:51ZengMDPI AGMedicina1010-660X1648-91442024-03-0160350110.3390/medicina60030501Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant <i>Enterococcus faecalis</i>, the Causative Agent of Dental AbscessAsmaa Ramadan0Mohamed O. Abdel-Monem1Noha K. El-Dougdoug2Alsayed E. Mekky3Shymaa A. Elaskary4Abdulaziz A. Al-Askar5Shimaa A Metwally6Ahmed F. El-Sayed7Gehad AbdElgayed8Ebrahim Saied9Mohamed Khedr10Botany and Microbiology Department, Faculty of Science, Benha University, Benha 13511, EgyptBotany and Microbiology Department, Faculty of Science, Benha University, Benha 13511, EgyptBotany and Microbiology Department, Faculty of Science, Benha University, Benha 13511, EgyptBotany and Microbiology Department, Faculty of Science, Al-Azhar University, Nasr, Cairo 11884, EgyptMedical Microbiology and Immunology Department, Faculty of Medicine, Menoufia University, Shibin El Kom 32511, EgyptBotany and Microbiology Department, Faculty of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi ArabiaMicrobiology and Immunology Department, Faculty of Pharmacy for Girls, Al-Azhar University, Cairo 4434004, EgyptMicrobial Genetics Department, Biotechnology Research Institute, National Research Centre, Giza 12622, EgyptIntegrated Molecular Plant Physiology Research, Department of Biology, University of Antwerp, 2020 Antwerp, BelgiumBotany and Microbiology Department, Faculty of Science, Al-Azhar University, Nasr, Cairo 11884, EgyptBotany and Microbiology Department, Faculty of Science, Al-Azhar University, Nasr, Cairo 11884, Egypt<i>Background and Objectives</i>: <i>Enterococcus faecalis (E. faecalis)</i> is a primary pathogen responsible for dental abscesses, which cause inflammation and pain when trapped between the crown and soft tissues of an erupted tooth. Therefore, this study aims to use specific phages as an alternative method instead of classical treatments based on antibiotics to destroy multidrug-resistant <i>E. faecalis</i> bacteria for treating dental issues. <i>Materials and Methods</i>: In the current study, twenty-five bacterial isolates were obtained from infected dental specimens; only five had the ability to grow on bile esculin agar, and among these five, only two were described to be extensive multidrug-resistant isolates. <i>Results</i>: Two bacterial isolates, <i>Enterococcus faecalis</i> A.R.A.01 [ON797462.1] and <i>Enterococcus faecalis</i> A.R.A.02, were identified biochemically and through 16S rDNA, which were used as hosts for isolating specific phages. Two isolated phages were characterized through TEM imaging, which indicated that <i>E. faecalis</i>_phage-01 had a long and flexible tail, belonging to the family <i>Siphoviridae</i>, while <i>E</i>. <i>faecalis</i>_phage-02 had a contractile tail, belonging to the family <i>Myoviridae.</i> Genetically, two phages were identified through the PCR amplification and sequencing of the RNA ligase of <i>Enterococcus</i> phage vB_EfaS_HEf13, through which our phages shared 97.2% similarity with <i>Enterococcus</i> phage vB-EfaS-HEf13 based on BLAST analysis. Furthermore, through in silico analysis and annotations of the two phages’ genomes, it was determined that a total of 69 open reading frames (ORFs) were found to be involved in various functions related to integration excision, replication recombination, repair, stability, and defense. In phage optimization, the two isolated phages exhibited a high specific host range with <i>Enterococcus faecalis</i> among six different bacterial hosts, where <i>E. faecalis</i>_phage-01 had a latent period of 30 min with 115.76 PFU/mL, while <i>E</i>. <i>faecalis</i>_phage-02 had a latent period of 25 min with 80.6 PFU/mL. They were also characterized with stability at wide ranges of pH (4–11) and temperature (10–60 °C), with a low cytotoxic effect on the oral epithelial cell line at different concentrations (1000–31.25 PFU/mL). <i>Conclusions</i>: The findings highlight the promise of phage therapy in dental medicine, offering a novel approach to combating antibiotic resistance and enhancing patient outcomes. Further research and clinical trials will be essential to fully understand the therapeutic potential and safety profile of these bacteriophages in human populations.https://www.mdpi.com/1648-9144/60/3/501bacterial dental abscesses<i>Enterococcus faecalis</i><i>E. faecalis</i>_phage-01<i>E. faecalis</i>_phage-02multi-drug resistanceoral epithelial cell line
spellingShingle Asmaa Ramadan
Mohamed O. Abdel-Monem
Noha K. El-Dougdoug
Alsayed E. Mekky
Shymaa A. Elaskary
Abdulaziz A. Al-Askar
Shimaa A Metwally
Ahmed F. El-Sayed
Gehad AbdElgayed
Ebrahim Saied
Mohamed Khedr
Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant <i>Enterococcus faecalis</i>, the Causative Agent of Dental Abscess
Medicina
bacterial dental abscesses
<i>Enterococcus faecalis</i>
<i>E. faecalis</i>_phage-01
<i>E. faecalis</i>_phage-02
multi-drug resistance
oral epithelial cell line
title Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant <i>Enterococcus faecalis</i>, the Causative Agent of Dental Abscess
title_full Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant <i>Enterococcus faecalis</i>, the Causative Agent of Dental Abscess
title_fullStr Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant <i>Enterococcus faecalis</i>, the Causative Agent of Dental Abscess
title_full_unstemmed Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant <i>Enterococcus faecalis</i>, the Causative Agent of Dental Abscess
title_short Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant <i>Enterococcus faecalis</i>, the Causative Agent of Dental Abscess
title_sort fully characterized effective bacteriophages specific against antibiotic resistant i enterococcus faecalis i the causative agent of dental abscess
topic bacterial dental abscesses
<i>Enterococcus faecalis</i>
<i>E. faecalis</i>_phage-01
<i>E. faecalis</i>_phage-02
multi-drug resistance
oral epithelial cell line
url https://www.mdpi.com/1648-9144/60/3/501
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