Abstract
In this study, we characterize three phages (SL1 SL2, and SL4), isolated from hospital sewage with lytic activity against clinical isolates of multi-drug resistant Pseudomonas aeruginosa (MDR-PA). The host spectrum ranged from 41% to 54%, with all three phages together covering 79% of all tested clinical isolates. Genome analysis revealed that SL1 (65,849 bp, 91 open reading frames ORFs) belongs to PB1-like viruses, SL2 (279,696 bp, 354 ORFs) to phiKZ-like viruses and SL4 (44,194 bp, 65 ORFs) to LUZ24-like viruses. Planktonic cells of four of five selected MDR-PA strains were suppressed by at least one phage with multiplicities of infection (MOIs) ranging from 1 to 10−6 for 16 h without apparent regrowth of bacterial populations. While SL2 was most potent in suppressing planktonic cultures the strongest anti-biofilm activity was observed with SL4. Phages were able to rescue bacteria-infected wax moth larvae (Galleria melonella) for 24 h, whereby highest survival rates (90%) were observed with SL1. Except for the biofilm experiments, the effect of a cocktail with all three phages was comparable to the action of the best phage alone; hence, there are no synergistic but also no antagonistic effects among phages. The use of a cocktail with these phages is therefore expedient for increasing host range and minimizing the development of phage resistance.
Highlights
Human phage therapy is currently being broadly reconsidered and multi-drug resistant organisms from the ESKAPE group (i.e., Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) are likely the prime candidates for which this treatment option might be realized in the Western world [1]
The interest in phage therapy is strong for P. aeruginosa, as this species is a notorious source of bacteremia in burn victims, urinary-tract infections in catheterized patients, and hospital-acquired pneumonia and respiratory infections in cystic fibrosis patients [2]
We describe the characterization of three selected phage isolates using current clinically relevant multi-drug resistant P. aeruginosa strains as target hosts
Summary
Human phage therapy is currently being broadly reconsidered and multi-drug resistant organisms from the ESKAPE group (i.e., Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) are likely the prime candidates for which this treatment option might be realized in the Western world [1]. The ubiquity and heterogeneity of P. aeruginosa has led to the evolution of distinct phage species that can be globally isolated from a variety of many different environmental niches [7,8]. This is in part reflected by the fact that among members of ESKAPE the number of available phages from official phage collection centers is highest for P. aeruginosa [9]. We describe the characterization of three selected phage isolates (each affiliated with a different phage genus, i.e., PB1-like, phiKZ-like and LUZ24-like viruses) using current clinically relevant multi-drug resistant P. aeruginosa strains as target hosts
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