Resistance, mechanism, and fitness cost of specific bacteriophages for Pseudomonas aeruginosa

Abstract

The bacteriophage is an effective adjunct to existing antibiotic therapy; however, in the course of bacteriophage therapy, host bacteria will develop resistance to bacteriophages, thus affecting the efficacy. Therefore, it is important to describe how bacteria evade bacteriophage attack and the consequences of the biological changes that accompany the development of bacteriophage resistance before the bacteriophage is applied. The specific bacteriophage vB3530 of Pseudomonas aeruginosa ( P. aeruginosa ) has stable biological characteristics, short incubation period, strong in vitro cleavage ability, and absence of virulence or resistance genes. Ten bacteriophage-resistant strains (TL3780-R) were induced using the secondary infection approach, and the plaque assay showed that vB3530 was less sensitive to TL3780-R. Identification of bacteriophage adsorption receptors showed that the bacterial surface polysaccharide was probably the adsorption receptor of vB3530. In contrast to the TL3780 parental strain, TL3780-R is characterized by the absence of long lipopolysaccharide chains, which may be caused by base insertion of wzy or deletion of galU . It is also intriguing to observe that, in comparison to the parent strain, the bacteriophage-resistant strains TL3780-R mostly exhibited a large cost of fitness (growth rate, biofilm formation, motility, and ability to produce enhanced pyocyanin). In addition, TL3780-R9 showed increased susceptibility to aminoglycosides and chlorhexidine, which may be connected to the loss and down-regulation of mexX expression. Consequently, these findings fully depicted the resistance mechanism of P. aeruginosa to vB3530 and the fitness cost of bacteriophage resistance, laying a foundation for further application of bacteriophage therapy. IMPORTANCE The bacteriophage is an effective adjunct to existing antibiotic therapy; However, bacteria also develop defensive mechanisms against bacteriophage attack. Thus, there is an urgent need to deeply understand the resistance mechanism of bacteria to bacteriophages and the fitness cost of bacteriophage resistance so as to lay the foundation for subsequent application of the phage. In this study, a specific bacteriophage vB3530 of P. aeruginosa had stable biological characteristics, short incubation period, strong in vitro cleavage ability, and absence of virulence or resistance genes. In addition, we found that P. aeruginosa may lead to phage resistance due to the deletion of galU and the base insertion of wzy , involved in the synthesis of lipopolysaccharides. Simultaneously, we showed the association with the biological state of the bacteria after bacteria acquire bacteriophage resistance, which is extremely relevant to guide the future application of therapeutic bacteriophages.

Using phage to drive selections toward restoring antibiotic sensitivity in Pseudomonas aeruginosa via chromosomal deletions

“…The results also suggest that LPS-targeting phages can also be used to indirectly induce a fitness trade-off via drug efflux pump function. Recently, Feng et al (2024) also demonstrated that resistance variants against a LPS-targeting phage exhibit extensive chromosomal deletions, including 290 genes such as galU, hmgA, mexX , and mexY . This deletion revealed increased susceptibility to aminoglycosides and chlorhexidine, likely due to the loss of mexX expression ( Feng et al, 2024 ).…”

“…Recently, Feng et al (2024) also demonstrated that resistance variants against a LPS-targeting phage exhibit extensive chromosomal deletions, including 290 genes such as galU, hmgA, mexX , and mexY . This deletion revealed increased susceptibility to aminoglycosides and chlorhexidine, likely due to the loss of mexX expression ( Feng et al, 2024 ). On the other hand, large chromosomal deletions containing mexX and mexY have also been reported to elicit resistance against ΦPIAS which is presumed to infect via the MexY as a receptor ( Koderi Valappil et al, 2021 ).…”