A study of antimicrobial effect of Lactiplantibacillus plantarum 8P-A3 supernatant on nosocomial isolates of Klebsiella pneumoniae and Acinetobacter baumannii | CMAC

A study of antimicrobial effect of Lactiplantibacillus plantarum 8P-A3 supernatant on nosocomial isolates of Klebsiella pneumoniae and Acinetobacter baumannii

Clinical Microbiology and Antimicrobial Chemotherapy. 2024; 26(4):505-513

​ Belova I.V., Shirokova I.Yu., Tochilina A.G., Kovalishena O.V., Belyanina N.A., Tulupov A.A., Molodtsova S.B., Seliverstov A.N., Kropotov V.S., Solovyova I.V.
10.36488/cmac.2024.4.505-513
wound infection Lactiplantibacillus plantarum 8P-A3 bacteriocins antimicrobial resistance K pneumoniae A baumannii
Section
Personal Experience
Type
Original Article
Publication
Clinical Microbiology and Antimicrobial Chemotherapy. 2024; 26(4):505-513

Objective.

To study antimicrobial effect of extracellular metabolites of probiotic L. plantarum 8P-A3 strain on antibiotic-resistant Klebsiella pneumoniae and Acinetobacter baumannii isolated from infected wounds.

Materials and Methods.

Bacteriological methods: semiquantitative sector method for extracting opportunistic microorganisms from an infected wound; a method for the identification of the antimicrobial activity of metabolites in a liquid milieu; The identification of bacteria was implemented with the help of the MALDI-TOF Autoflex mass spectrometer (Bruker Daltonics, Germany). Antimicrobial susceptibility testing was performed using SensiLaTest test systems (Erba Lachema, Czech Republic), analyzer Vitek 2 and disk-diffusion method. The prevalence of resistance genes was determined using BakResista GLA reagent kits by real-time PCR. In order to obtain L. plantarum 8P-A3 supernatant, cells were precipitated by centrifugation and then filtered using a membrane (Merck Millipore, Germany). Metabolomic profiling of the supernatant was implemented by HPLC/MS on mass spectrometer LCMS-8050 (Shimadzu, Japan). Whole-genome sequencing was performed on MiSeq platform (Illumina Inc., USA), and data processing was conducted using Trimmomatic, Spades, Prokka, RAST and Bagel 4 software tools.

Results.

All strains of A. baumannii and K. pneumoniae are multidrug-resistant and contain a range of beta-lactamases and carbapenemase in their genome. The results of metabolomic profiling showed that L. plantarum 8P-A3 supernatant contains a significant amount of lactic, valeric, isovaleric, succinic and acetic acids. Analysis of L. plantarum 8P-A3 genome found key enzymes for synthesis of lactic and acetic acids, hydrogen peroxide, lysozyme and plantaricins A, EF and NC8. This experiment proved that the supernatant completely inhibits the growth of nosocomial isolates of A. baumannii and K. pneumoniae.

Conclusions.

Using whole-genome sequencing, the ability of L. plantarum 8P-A3 strain to synthesize extracellular metabolites with antimicrobial activity was determined. Metabolomic profiling of its supernatant proved the presence of high concentrations of organic acids in it. The presence of all these metabolites determined significant antimicrobial effect of the supernatant on multidrug-resistant nosocomial K. pneumoniae and A. baumannii isolates and showed potential for use as an adjunctive treatment of infected wounds.

Belova I.V.

Privolzhsky Research Medical University, Nizhny Novgorod, Russia
Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology, Nizhny Novgorod, Russia

Shirokova I.Yu.

Privolzhsky Research Medical University, Nizhny Novgorod, Russia

Anna G. Tochilina

Privolzhsky Research Medical University, Nizhny Novgorod, Russia
Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology, Nizhny Novgorod, Russia

Kovalishena O.V.

Privolzhsky Research Medical University, Nizhny Novgorod, Russia

Belyanina N.A.

Privolzhsky Research Medical University, Nizhny Novgorod, Russia

Alexander A. Tulupov

Privolzhsky Research Medical University, Nizhny Novgorod, Russia

Molodtsova S.B.

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology, Nizhny Novgorod, Russia

Seliverstov A.N.

Privolzhsky Research Medical University, Nizhny Novgorod, Russia

Kropotov V.S.

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology, Nizhny Novgorod, Russia

Solovyova I.V.

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology, Nizhny Novgorod, Russia

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