Approaches to Improving Methods of Isolation and Identification of Pseudomonas aeruginosa

Introduction: According to the World Health Organization, Pseudomonas aeruginosa is a high-priority microorganism due to its acquired antibiotic resistance and the ability to produce many virulence factors. Current regulatory documents and guidelines define the algorithm of Pseudomonas aeruginosa detection during bacteriological testing of objects. The laboratory prepared Blesk medium is of preference.

Objectives: To improve methods of P. aeruginosa isolation and identification (i.e. to increase the efficiency of the Pseudomonas aeruginosa isolation procedure) by developing an optimal composition of the modified standard dry domestic nutrient medium for selective isolation of P. aeruginosa (Blesk (“Shine”) medium).

Materials and Methods: We analyzed regulatory documents and guidelines on the use of nutrient media to detect P. aeruginosa and then tested dry nutrient media and the media produced in the laboratory of the State Research Center for Applied Microbiology and Biotechnology. Sensitivity, selective and differentiating properties of the media were evaluated using a set of reference strains of microorganisms provided by the State Collection of Pathogenic Microorganisms and Cell Cultures (SCPM-Obolensk).

Results: A dry modified domestic nutrient medium for selective isolation of P. aeruginosa (Blesk medium) has been developed and put into production. A unique protein base (casein pancreatic hydrolysate dried in the presence of Tween, modified) has been created that enhances the sensitivity of the medium and intensifies the specific golden shine of bright red colonies, which is an important diagnostic feature for pseudomonads growing on this medium. Optimal combinations of the main components when developing the Blesk nutrient medium fostered the growth of all P. aeruginosa reference strains and inhibited the growth of Gram-positive and most Gram-negative bacteria, including TTX-resistant strains.

Conclusion: Our findings proved that the balanced composition of the Blesk medium ensures the growth and detection of low concentrations of P. aeruginosa and prevents the growth of associative microbes, which is a significant advantage when testing the samples highly contaminated with foreign microflora and containing few pathogenic cells.

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