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Quantifying the Effects of Antimicrobials on In vitro Biofilm Architecture using COMSTAT Software
Published on: December 14, 2020
Effects of аntimicrobials on Pseudomonas aeruginosa biofilm formation
U M Nemchenko1, K O Sitnikova1, N L Belkova1
1Scientific Сentre for Family Health and Human Reproduction Problems, Irkutsk, Russia.
Abstract:
Pseudomonas aeruginosa is one of the most problematic pathogens in medical institutions, which may be due to the ability of this microorganism to exist in a biofilm, which increases its resistance to antimicrobials, as well as its prevalence and survival ability in the external environment. This work aimed to evaluate the antimicrobial susceptibility of P. aeruginosa strains in planktonic and biofilm forms. We studied 20 strains of P. aeruginosa collected during 2018-2021 by specialists from the Laboratory of Microbiome and Microecology of the Scientific Centre for Family Health and Human Reproduction Problems. The identification of strains was carried out using test systems for differentiating gram-negative non-fermenting bacteria (NEFERMtest 24 Erba Lachema s.r.o., Czech Republic), and confirmed by mass spectrometric analysis and 16S rRNA gene sequencing. Antimicrobial activity was assessed by the degree of inhibition of cell growth in planktonic and biofilm forms (on a flat-bottomed 96-well plastic immunological plate). All clinical isolates of P. aeruginosa were biofilm formers, 47.6 % of the isolates were weak biofilm formers, and 52.4 % of the isolates were moderate biofilm formers. Planktonic cells and the forming biofilm of the tested P. aeruginosa strains were carbapenems-resistant. Biofilm formation was suppressed in more than 90 % of cases by the agents of the cephalosporin and aminoglycoside groups. Antimicrobial susceptibility of P. aeruginosa strains in the formed biofilm was significantly lower (p < 0.05). Carbapenems and cephalosporins did not affect the mature biofilms of the tested P. aeruginosa strains in more than 60 % of cases. Only non-beta-lactam antibiotics (ciprofloxacin and amikacin) suppressed the growth of planktonic cells and destroyed the mature biofilm. The revealed differences in the effect of the tested antimicrobials on the P. aeruginosa strains biofilms correlate with resistance to a number of antibiotics. To prevent biofilm formation in the hospital strains of P. aeruginosa, the use of ceftazidime may be recommended, and antimicrobials such as ciprofloxacin and amikacin may be used to affect mature biofilms of P. aeruginosa.
Insights
Pseudomonas aeruginosa biofilms are resistant to carbapenems and cephalosporins. Ciprofloxacin and amikacin effectively target both planktonic cells and mature biofilms of this problematic pathogen.
Area of Science:
- Medical Microbiology
- Antimicrobial Resistance
- Biofilm Formation
Background:
- Pseudomonas aeruginosa is a major healthcare-associated pathogen.
- Its ability to form biofilms significantly increases antimicrobial resistance.
- Understanding susceptibility in different growth forms is crucial for effective treatment.
Purpose of the Study:
- To evaluate the antimicrobial susceptibility of Pseudomonas aeruginosa strains in both planktonic and biofilm forms.
- To compare the efficacy of different antibiotic classes against P. aeruginosa biofilms.
- To identify potential therapeutic strategies against resistant strains.
Main Methods:
- Studied 20 P. aeruginosa clinical isolates (2018-2021).
- Identified strains using NEFERMtest 24, mass spectrometry, and 16S rRNA sequencing.
- Assessed antimicrobial activity by measuring growth inhibition in planktonic and biofilm cultures using 96-well plates.
Main Results:
- All tested P. aeruginosa isolates formed biofilms (weak and moderate).
- Planktonic cells and forming biofilms showed carbapenem resistance.
- Cephalosporins and aminoglycosides suppressed biofilm formation (>90%), but were less effective against mature biofilms (>60% resistance).
- Ciprofloxacin and amikacin were effective against both planktonic cells and mature biofilms.
Conclusions:
- Pseudomonas aeruginosa clinical isolates are significant biofilm formers with high carbapenem resistance.
- Mature biofilms exhibit reduced susceptibility to carbapenems and cephalosporins.
- Ceftazidime may prevent biofilm formation, while ciprofloxacin and amikacin are recommended for treating mature P. aeruginosa biofilms.
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