In Vivo Application of Bacteriophage as a Potential Therapeutic Agent To Control OXA-66-Like Carbapenemase-Producing Acinetobacter baumannii Strains Belonging to Sequence Type 357

Abstract

The increasing prevalence of carbapenem-resistant Acinetobacter baumannii (CRAB) strains in intensive care units has caused major problems in public health worldwide. Our aim was to determine whether this phage could be used as an alternative therapeutic agent against multidrug-resistant bacterial strains, specifically CRAB clinical isolates, using a mouse model. Ten bacteriophages that caused lysis in CRAB strains, including bla OXA-66-like genes, were isolated. YMC13/01/C62 ABA BP (phage B-C62), which showed the strongest lysis activity, was chosen for further study by transmission electron microscopy (TEM), host range test, one-step growth and phage adsorption rate, thermal and pH stability, bacteriolytic activity test, genome sequencing and bioinformatics analysis, and therapeutic effect of phage using a mouse intranasal infection model. The phage B-C62 displayed high stability at various temperatures and pH values and strong cell lysis activity in vitro. The phage B-C62 genome has a double-stranded linear DNA with a length of 44,844 bp, and known virulence genes were not identified in silico. In vivo study showed that all mice treated with phage B-C62 survived after intranasal bacterial challenge. Bacterial clearance in the lung was observed within 3 days after bacterial challenge, and histologic damage also improved significantly; moreover, no side effects were observed. IMPORTANCEIn our study, the novel A. baumannii phage B-C62 was characterized and evaluated in vitro, in silico, and in vivo. These results, including strong lytic activities and the improvement of survival rates, showed the therapeutic potential of the phage B-C62 as an antimicrobial agent. This study reports the potential of a novel phage as a therapeutic candidate or nontoxic disinfectant against CRAB clinical isolates in vitro and in vivo. B acteriophages (phages) are viruses that infect bacteria and are ubiquitous in the biosphere (1). Recently, phages have been reported as therapeutic agents to treat bacterial infectious diseases in humans and animals (2-5). Phages possess several advantages as therapeutic agents over conventional antibiotics, in that they have precise effective bacteriolytic activity against target bacteria and have shown no critical side effects to date (6-8).Antimicrobials have been crucial for the prevention and treatment of infectious diseases in humans, animals, and plants since the 1940s (9). Today, however, certain isolated strains have exhibited resistance to all currently available commercial antimicrobial agents (9, 10). Many scientists and medical professionals have emphasized the urgent need to prevent the emergence and spread of drug-resistant bacteria (11). Despite efforts to reduce the dissemination of drug-resistant pathogens in health care settings, antimicrobial resistance has been constantly increasing and becoming a serious global clinical challenge with little accompanying novel antibiotic discovery (12). Hence, phages are currently being viewed as alternative therapeutic agents (6).Acine...

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