Cell-free supernatant of probiotic bacteria exerted antibiofilm and antibacterial activities against Pseudomonas aeruginosa: A novel biotic therapy

Mariana Martins Drumond; Ana Paula Tapia-Costa; Elisabeth Neumann; Álvaro Cantini Nunes; Jorge Wanderson Barbosa; Diego E Kassuha; Pamela Mancha-Agresti

doi:10.3389/fphar.2023.1152588

Cell-free supernatant of probiotic bacteria exerted antibiofilm and antibacterial activities against Pseudomonas aeruginosa: A novel biotic therapy

Front Pharmacol. 2023 Jun 15:14:1152588. doi: 10.3389/fphar.2023.1152588. eCollection 2023.

Authors

Mariana Martins Drumond^{1

2}, Ana Paula Tapia-Costa³, Elisabeth Neumann⁴, Álvaro Cantini Nunes⁵, Jorge Wanderson Barbosa⁶, Diego E Kassuha³, Pamela Mancha-Agresti²

Affiliations

¹ Centro Federal de Educação Tecnológica de Minas Gerais (CEFET/MG), Departamento de Ciências Biológicas, Belo Horizonte, Minas Gerais, Brazil.
² Centro Federal de Educação Tecnológica de Minas Gerais (CEFET/MG), Programa de Pós Graduação em Engenharia de Materiais, Belo Horizonte, Minas Gerais, Brazil.
³ Instituto de Investigaciones en Ciencias Químicas, Facultad de Ciencias Químicas y Tecnológicas, Universidad Católica de Cuyo, San Juan, Argentina.
⁴ Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
⁵ Laboratório de Genética Molecular de Protozoários Parasitas, Departamento de Genética, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Minas Gerais, Brazil.
⁶ Centro Federal de Educação Tecnológica de Minas Gerais (CEFET/MG), Departamento de Engenharia de Materiais, Belo Horizonte, Minas Gerais, Brazil.

Abstract

Aim: This study aims to verify the antibacterial and antibiofilm action of cell-free spent medium (CFSM) from four lactic acid bacteria with potential probiotic characteristics (Lactiplantibacillus plantarum, Lactobacillus acidophilus, Lactobacillus johnsonii, and Lactobacillus delbrueckii) against two Pseudomonas aeruginosa strains. Main methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the CFSM, antibacterial activity by analysing the formation of inhibition zones, and inhibition of planktonic cultures were determined. Whether an increase in the concentration of CFSM influenced the growth of pathogenic strains and the anti-adhesive activity of the CFSM in biofilm formation (crystal violet and MTT assays) were determined, which were all corroborated by using scanning electron microscopy. Key findings: The relationship between the MIC and MBC values showed a bactericidal or bacteriostatic effect for all the cell-free spent media (CFSMs) tested for P. aeruginosa 9027™ and 27853™ strains. The CFSM supplemental doses of 18 or 22%, 20 or 22%, 46 or 48%, and 50 or 54% of L. acidophilus, L. delbrueckii, L. plantarum, and L. johnsonii, respectively, could completely inhibit the growth of both pathogen strains. The antibiofilm activity of the CFSM in three biofilm conditions (pre-coated, co-incubated, and preformed) demonstrated values ranging between 40% and 80% for biofilm inhibition, and similar results were observed for cell viability. Significance: This work provides strong evidence that the postbiotic derived from different Lactobacilli could be practical as an adjuvant therapy for reducing the use of antibiotics, being a good candidate to overcome the growing challenge of hospital infections due to this pathogen.

Keywords: Pseudomonas aeruginosa; biofilm; lactic acid bacteria; postbiotics fraction (supernatant); skin pathogens.

Grants and funding

This research was funded by Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)—grant number APQ-00766-21, Brasil, and by the Rede Mineira de Engenharia de Tecidos e Terapia Celular, FAPEMIG—grant number RED-00570/16.