Bacteriophage as a potential therapy to control antibiotic-resistant Pseudomonas aeruginosa infection through topical application onto a full-thickness wound in a rat model

Abstract

Background Antibiotic-resistant Pseudomonas aeruginosa (P. aeruginosa) is one of the most critical pathogens in wound infections, causing high mortality and morbidity in severe cases. However, bacteriophage therapy is a potential alternative to antibiotics against P. aeruginosa. Therefore, this study aimed to isolate a novel phage targeting P. aeruginosa and examine its efficacy in vitro and in vivo. Results The morphometric and genomic analyses revealed that ZCPA1 belongs to the Siphoviridae family and could infect 58% of the tested antibiotic-resistant P. aeruginosa clinical isolates. The phage ZCPA1 exhibited thermal stability at 37 °C, and then, it decreased gradually at 50 °C and 60 °C. At the same time, it dropped significantly at 70 °C, and the phage was undetectable at 80 °C. Moreover, the phage ZCPA1 exhibited no significant titer reduction at a wide range of pH values (4–10) with maximum activity at pH 7. In addition, it was stable for 45 min under UV light with one log reduction after 1 h. Also, it displayed significant lytic activity and biofilm elimination against P. aeruginosa by inhibiting bacterial growth in vitro in a dose-dependent pattern with a complete reduction of the bacterial growth at a multiplicity of infection (MOI) of 100. In addition, P. aeruginosa-infected wounds treated with phages displayed 100% wound closure with a high quality of regenerated skin compared to the untreated and gentamicin-treated groups due to the complete elimination of bacterial infection. Conclusion The phage ZCPA1 exhibited high lytic activity against MDR P. aeruginosa planktonic and biofilms. In addition, phage ZCPA1 showed complete wound healing in the rat model. Hence, this research demonstrates the potential of phage therapy as a promising alternative in treating MDR P. aeruginosa.

Topically applied bacteriophage to control multi-drug resistant Pseudomonas aeruginosa-infected wounds in a New Zealand rabbit model

“…All New Zealand rabbits treated with the phage cocktail in this study exhibited epidermal re-epithelialization and regeneration of skin appendages, similar to New Zealand rabbits before wound infection. In contrast, ceftriaxone-treated New Zealand rabbits had incomplete epidermal formation and lacked skin appendages, similar to the findings of ( Rezk et al, 2022 ). Based on photographic records of wound healing and histopathological analysis, this study demonstrates the potential of phages to treat drug-resistant infections.…”

“…Regarding histopathological results, accelerated skin healing and remodeling were observed in the phage cocktail treatment group, which was equivalent to the wound healing in the negative control group. Skin regeneration, an indicator for assessing the efficacy of wound treatment, re-epithelization is critical to successful wound healing ( Safferling et al, 2013 ; Rezk et al, 2022 ). All New Zealand rabbits treated with the phage cocktail in this study exhibited epidermal re-epithelialization and regeneration of skin appendages, similar to New Zealand rabbits before wound infection.…”

Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2

“…The host range of phage ZCKP2 was determined against 30 clinical isolates of MDR K . pneumoniae by using the spot assay in triplicate as previously mentioned [ 48 , 53 ]. Briefly, 100 µl of freshly prepared culture of each bacterial strain was added to 4 mL of 0.5% top agar, which was subsequently poured onto a base TSA agar.…”

“…Shortly, host strain KP/08 was grown at 37 °C to exponential phase (~ 10 8 CFU/mL) and incubated with ZCKP2 phage (~ 10 7 PFU/mL) at an MOI of 0.1. Aliquots of the infected culture were serially diluted at each time point to count the number of phages present using a spotting test [ 53 ]. Two aliquots were collected at each time point, the first one was treated with 1% (v/v) chloroform to induce the release of the intracellular phage, while the second one was spotted without chloroform to determine the phage infective center (IC).…”

Pseudomonas aeruginosa PAO1 outer membrane vesicles-diphtheria toxoid conjugate as a vaccine candidate in a murine burn model

“…Vaccines have also been shown to significantly reduce antibiotic prescription and thus reduce antimicrobial resistance 9 . Infection of a burnt wound by antibiotic-resistant organisms, such as multidrug-resistant (MDR) and extensively drug-resistant (XDR) P. aeruginosa , has been reported as the leading cause of mortality and morbidity 10 . Remarkably, these strains possess considerable ability to survive in healthcare settings, particularly in burn care units.…”