Antimicrobial resistance, virulence factors, and genotypes of Pseudomonas aeruginosa clinical isolates from Gorgan, northern Iran.

Pseudomonas aeruginosa is an important nosocomial pathogen with a capacity of resistance to multiple antibiotics and production of various extracellular and cell-associated virulence factors that clearly contribute to its pathogenicity. The objective of this study was to investigate the antibiotic susceptibility, virulence factors, and clonal relationship among clinical isolates of P. aeruginosa. Different clinical specimens from hospitalized patients were investigated for P. aeruginosa. Susceptibility of the isolates was evaluated by disc diffusion and broth microdilution methods, as described by the Clinical and Laboratory Standards Institute (CLSI) guideline. A total of 97 P. aeruginosa isolates were recovered from clinical specimens. The percentage of isolates resistant to antimicrobials was imipenem 25.77%, meropenem 15.46%, gentamicin 16.49%, tobramycin 15.46%, amikacin 16.49%, ciprofloxacin 20.61%, levofloxacin 24.74, ceftazidime 20.61%, piperacillin 15.46%, piperacillin/tazobactam 12.37%, colistin 9.27%, and polymyxin B 11.34%. Of isolates, 87.62% possessed ß-hemolytic activity, 78.35% lecithinase, 59.8% elastase, 37.11% DNase, and 28.86% twitching motility. The frequency of virulence genes in isolates was lasB 82.47%, plcH 82.47%, exoA 58.76%, exoS 56.7%, and pilA 10.3%. ERIC-PCR typing clustered P. aeruginosa isolates to 19 common types (CT1-CT19) containing isolates from different hospitals and 43 single types (ST1-ST43). Colistin and polymyxin B were the most effective agents against the majority of P. aeruginosa isolates, emphasizing the effort to maintain their antibacterial activity as last-line therapy. The frequency of some virulence factors and genes was noticeably high, which is alarming. In addition, more effective strategies and surveillance are necessary to confine and prevent the inter-hospital and/or intra-hospital dissemination of P. aeruginosa between therapeutic centers.

In vitro activities of cellulase and ceftazidime, alone and in combination against Pseudomonas aeruginosa biofilms.

BACKGROUND: Biofilms are a main pathogenicity feature of Pseudomonas aeruginosa and has a significant role in antibiotic resistance and persistent infections in humans. We investigated the in vitro activities of antibiotic ceftazidime and enzyme cellulase, either alone or in combination against biofilms of P. aeruginosa. RESULTS: Both ceftazidime and cellulase significantly decreased biofilm formation in all strains in a dose-dependent manner. Combination of enzyme at concentrations of 1.25, 2.5, 5, and 10 U/mL tested with 1/16× MIC of antibiotic led to a significant reduction in biofilm biomass. Cellulase showed a significant detachment effect on biofilms at three concentrations of 10 U/mL, 5 U/mL, and 2.5 U/mL. The MIC, MBC, and MBEC values of ceftazidime were 2 to 4 µg/mL, 4 to 8 µg/mL, and 2048 to 8192 µg/mL. When combined with cellulase, the MBECs of antibiotic showed a significant decrease from 32- to 128-fold. CONCLUSIONS: Combination of the ceftazidime and the cellulase had significant anti-biofilm effects, including inhibition of biofilm formation and biofilm eradication in P. aeruginosa. These data suggest that glycoside hydrolase therapy as a novel strategy has the potential to enhance the efficacy of antibiotics and helps to resolve biofilm-associated wound infections caused by this pathogen.

Evaluation of antimicrobial resistance, biofilm forming potential, and the presence of biofilm-related genes among clinical isolates of Pseudomonas aeruginosa.

OBJECTIVES: Pseudomonas aeruginosa is known as a leading cause of nosocomial infections worldwide. Antimicrobial resistance and biofilm production, as two main virulence factors of P. aeruginosa, are responsible for the persistence of prolonged infections. In this study, antimicrobial susceptibility pattern and phenotypic and genotypic characteristics of biofilm of P. aeruginosa were investigated. RESULTS: A total of 80 clinical P. aeruginosa isolates were obtained. Isolates showed resistance to all antibiotics with a rate from 12.5% (n = 10) against amikacin and piperacillin/tazobactam to 23.75% (n = 19) to levofloxacin. Multidrug-resistant P. aeruginosa accounted for 20% (n = 16). 83.75% (n = 67) of isolates showed biofilm phenotype. All three biofilm-related genes were found simultaneously in 87.5% (n = 70) of P. aeruginosa and 13.5% (n = 10) of the isolates had none of the genes tested. From the results of the present study, combination therapy including an anti-pseudomonal beta-lactam (piperacillin/tazobactam or ceftazidime) and an aminoglycoside or carbapenems (imipenem, meropenem) with fluoroquinolones in conjunction with an aminoglycoside can be used against Pseudomonas infections. However, reasonable antimicrobial use and high standards of infection prevention and control are essential to prevent further development of antimicrobial resistance. Combination strategies based on the proper anti-pseudomonal antibiotics along with anti-biofilm agents can also be selected to eradicate biofilm-associated infections.

Heteroresistance to colistin in oxacillinase-producing carbapenem-resistant Acinetobacter baumannii clinical isolates from Gorgan, Northern Iran.

OBJECTIVES: Colistin resistance rates are rising globally among multidrug-resistant Gram-negative bacilli, including Acinetobacter baumannii (A. baumannii). A new type of resistance - heteroresistance - has also been reported to colistin in clinical A. baumannii isolates. This study investigated the presence of colistin heteroresistance in carbapenem-resistant A. baumannii clinical isolates. METHODS: Different clinical specimens from hospitalised patients were investigated for A. baumannii. The MICs to imipenem, meropenem and colistin were determined by broth microdilution. PCR was performed to detect OXA-type carbapenemase genes (blaOXA-23-like, blaOXA-24/40-like, blaOXA-51-like, blaOXA-58-like, and blaOXA-143-like). Heteroresistance to colistin was examined using the population analysis profiles method. Genotypic relatedness of the isolates was analysed by enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR). RESULTS: Overall, 71 A. baumannii isolates were recovered from clinical specimens. Of these, 27 (38.03%) and 44 (61.97%) isolates were carbapenem-susceptible and carbapenem-resistant, respectively. In addition, 67 (94.36%) isolates were susceptible to colistin, with MICs between 0.25-2 µg/mL. Among the 44 selected carbapenem-resistant colistin-susceptible isolates, the frequency of blaOXA-51-like, blaOXA-23-like and blaOXA-24/40-like genes was 100%, 77.27% and 43.18%, respectively. Nine of 44 (20.45%) isolates were characterised as colistin-heteroresistant with subpopulations growing at 6-8 µg/mL, whereas two of 44 (4.54%) presented heterogeneous subpopulations growing at up to 1 µg/mL of colistin. ERIC­PCR typing clustered A. baumannii isolates to 10 common types (CT1-CT10) containing isolates from different hospitals and 12 single types (ST1-ST12). CONCLUSIONS: A. baumannii with a colistin heteroresistance phenotype was common. This could be of great concern since colistin is often used as a last-resort drug for treating A. baumannii infections, highlighting that care is necessary with colistin monotherapy. In addition, more effective strategies and surveillance are required to confine and prevent the inter-hospital and/or intra-hospital dissemination of A. baumannii between therapeutic centres.