Background: Methicillin-resistant Staphylococcus aureus (MRSA) is an important zoonotic pathogen worldwide. Infections due to MRSA are associated with higher mortality rates compared with methicillin-susceptible S. aureus . Meanwhile, bacteriophages have been shown to overcome the emergence of MRSA.
Methods: Phage PHB22a, PHB25a, PHB38a, and PHB40a were isolated. Here, we evaluated the ability of a phage cocktail containing phages PHB22a, PHB25a, PHB38a, and PHB40a against MRSA S-18 strain in vivo and in vitro. Phage whole-genome sequencing, host-range determination, lytic activity, and biofilm clearance experiments were performed in vitro. Galleria mellonella larvae and a mouse systemic infection model to evaluate the efficacy of phage therapy in vivo.
Results: The phage cocktail exhibited enhanced antibacterial and anti-biofilm effects compared to the single phage. Phage cocktail contained with Ca²⁺/Zn²⁺ significantly reduced the number of viable bacteria (24-h or 48-h biofilm) by more than 0.81-log compared to the phage cocktail alone. Furthermore, we demonstrated that the addition of Ca²⁺ and Zn²⁺ phage cocktail could increase the survival rate of G. mellonella larvae infected with S. aureus by 10% compared with phage cocktail alone. This was further confirmed in the mouse model, which showed a 2.64-log reduction of host bacteria S-18, when Ca²⁺ and Zn²⁺ were included in the cocktail compared with the phage cocktail alone.
Conclusion: Our results indicated that phage cocktail supplemented with Ca²⁺/Zn²⁺ could effectively remove bacteria in biofilms and mice tissues infected with S. aureus .
Keywords: methicillin-resistant Staphylococcus aureus , phage cocktail, metal ions, biofilm, phage therapy