Whole genome sequencing of resistance and virulence genes in multi-drug resistant Pseudomonas aeruginosa.

BACKGROUND: Pseudomonas aeruginosa is an opportunistic bacterium that causes serious hospital-acquired infections. To assess the risk of clinically isolated P. aeruginosa to human health, we analyzed the resistance and virulence mechanisms of a collection of clinical isolates. METHODS: This was a retrospective study in which P. aeruginosa isolates collected from January 1, 2018 to August 31, 2019 were analyzed using phenotypic and whole-genome sequencing (WGS) methods. The analysis included 48 clinical samples. Median patient age was 54.0 (29.5) years, and 58.3% of patients were women. Data from the microbiology laboratory database were reviewed to identify P. aeruginosa isolates. All unique isolates available for further testing were included, and related clinical data were collected. Infections were defined as hospital acquired if the index culture was obtained at least 48 h after hospitalization. RESULTS: High-risk P. aeruginosa clones, including sequence types (STs) ST235 and ST111, were identified, in addition to 12 new STs. The isolates showed varying degrees of biofilm formation ability when evaluated at room temperature, along with reduced metabolic activity, as measured by metabolic staining, suggesting their ability to evade antimicrobial therapy. Most isolates (77.1%) were multidrug resistant (MDR), with the highest resistance and susceptibility rates to beta-lactams and colistimethate sodium, respectively. CONCLUSIONS: The MDR phenotypes of the examined isolates can be explained by the high prevalence of efflux-mediated resistance- and hydrolytic enzyme-encoding genes. These isolates had high cytotoxic potential, as indicated by the detection of toxin production-related genes.

Evaluation of riboflavin and ultraviolet light treatment against Klebsiella pneumoniae in whole blood-derived platelets: A pilot study.

BACKGROUND: Bacterial contamination of platelet concentrates (PCs) is the predominant cause of infectious transfusion reactions. The Pathogen Inactivation Mirasol system was implemented at the King Faisal Specialist Hospital (Saudi Arabia) to reduce the risk of transfusing contaminated PCs. This pilot study evaluated the effectiveness of Mirasol against Klebsiella pneumoniae, a pathogen associated with transfusion reactions, in whole blood-derived PCs. STUDY DESIGN AND METHODS: Whole blood (WB) units inoculated with one of six K. pneumoniae strains (five clinical isolates and ATCC-700603) at a concentration of 3-38 CFU/unit, were processed using the platelet-rich plasma (PRP) method. Each spiked PC was pooled with four unspiked units. The pooled PC was split into three Mirasol storage bags: an untreated unit (control), and two units treated with Mirasol at 26 and 32 h post-WB collection, respectively. PC samples obtained before and after Mirasol treatment were used for BacT/ALERT cultures and determination of bacteria quantification. Each experiment was repeated three independent times. RESULTS: Five strains were detected prior to PC treatment (24 h post-WB spiking), while one clinical isolate was not detected. Mirasol treatment after 26 h of WB collection resulted in complete inactivation of all K. pneumoniae strains. However, treatment 32 h post-WB collection resulted in the breakthrough of one clinical isolate in two of the three replicates with ~7.8 log10 CFU/unit detected on day 5 of PC storage. CONCLUSION: Delayed Mirasol treatment from 26 to 32 h post-WB collection, resulted in one breakthrough. These results highlight the importance of minimizing the time between WB collection and PI treatment.