Topically Applied Bacteriophage to Control Multi-Drug Resistant Klebsiella pneumoniae Infected Wound in a Rat Model.

Mohamed S Fayez,Nouran Rezk,Mona M Agwa,Abdallah S Abdelsattar,Toka A Hakim,Nada N Montaser,Ayman El-Shibiny,Mohamed Abdelmoteleb,Rania G Aly

doi:10.3390/antibiotics10091048

Abstract

(Background): Multi-drug-resistant Klebsiella pneumoniae (MDR-KP) has steadily grown beyond antibiotic control. Wound infection kills many patients each year, due to the entry of multi-drug resistant (MDR) bacterial pathogens into the skin gaps. However, a bacteriophage (phage) is considered to be a potential antibiotic alternative for treating bacterial infections. This research aims at isolating and characterizing a specific phage and evaluate its topical activity against MDR-KP isolated from infected wounds. (Methods): A lytic phage ZCKP8 was isolated by using a clinical isolate KP/15 as a host strain then characterized. Additionally, phage was assessed for its in vitro host range, temperature, ultraviolet (UV), and pH sensitivity. The therapeutic efficiency of phage suspension and a phage-impeded gel vehicle were assessed in vivo against a K. pneumoniae infected wound on a rat model. (Result): The phage produced a clear plaque and was classified as Siphoviridae. The phage inhibited KP/15 growth in vitro in a dose-dependent pattern and it was found to resist high temperature (˂70 °C) and was primarily active at pH 5; moreover, it showed UV stability for 45 min. Phage-treated K. pneumoniae inoculated wounds showed the highest healing efficiency by lowering the infection. The quality of the regenerated skin was evidenced via histological examination compared to the untreated control group. (Conclusions): This research represents the evidence of effective phage therapy against MDR-KP.

Highlights

Klebsiella pneumoniae (K. pneumoniae), is a non-motile, encapsulated, Gram-negative bacteria that belongs to the Enterobacteriaceae family [1]
The Polymerase Chain Reaction (PCR) was confirmed that the twenty isolated K. pneumonia, including KP/15 strain by a 130 bp band corresponding to the conserved region in 16S-23S internal transcribed spacer region of K. pneumoniae
The therapeutic efficiency of phage ZCKP8 was examined on rats to treat full-thickness wounds infected with K. pneumoniae clinical isolate (KP/15) which is resistant to multiple antibiotics, including Amikacin (AK; 30 μg), Aztreonam (ATM 30 μg), Ciprofloxacin (CIP; 5 μg), Ceftazidime (CAZ; 30 μg), Cefepime (Fep; 30 μg), Gentamicin (CN; 10 μg), Imipenem (IPM; 10 μg), Levofloxacin (LEV; 5 μg), Sulfamethoxazole/trimethoprim (SXT; 25 μg), Tigecycline (TGC; 15 μg), piperacillin/tazobactam (TZP; 110 μg), and Linezolid (LZD; 30 μg)

Summary

Introduction

Klebsiella pneumoniae (K. pneumoniae), is a non-motile, encapsulated, Gram-negative bacteria that belongs to the Enterobacteriaceae family [1]. It is mainly responsible for primary severe community-onset infections, including hepatic abscess, necrotizing pneumonia, and nosocomial infections of the urinary tract, respiratory tract, wounds, and bloodstream [2]. K. pneumoniae infections are often considered one of the most fatal life-threatening infections to infants, older people, and immunocompromised patients as they are linked to both high morbidity and mortality rates [4]. The World Health Organization (WHO) estimates that drug-resistant bacterial infections could kill approximately 10 million people per year by 2050 [8]

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