Intest Res.  2019 Apr;17(2):192-201. 10.5217/ir.2018.00106.

Evaluation of commercial probiotic lactic cultures against biofilm formation by Cronobacter sakazakii

• Jamwal A
• Sharma K
• Chauhan R
• Bansal S
• Goel G

• KMID: 2449960
• DOI: http://doi.org/10.5217/ir.2018.00106

Abstract

BACKGROUND/AIMS
Cronobacter sakazakii, an emergent pathogen is considered as a major concern to infants and neonates fed on reconstituted powdered infant milk formula. In conjunction with many other factors, biofilm forming capacity adds to its pathogenic potential. In view of the facts that infants are at highest risk to C. sakazakii infections, and emerging antibiotic resistance among pathogens, it is imperative to evaluate probiotic cultures for their efficacy against C. sakazakii. Therefore, pure probiotic strains were isolated from commercial probiotic products and tested for their antimicrobial and anti-biofilm activities against C. sakazakii.
METHODS
A total of 6 probiotic strains were tested for their antibiotic susceptibility followed by antimicrobial activity using cell-free supernatant (CFS) against C. sakazakii. The inhibitory activity of CFS against biofilm formation by C. sakazakii was determined using standard crystal violet assay and microscopic observations.
RESULTS
All the probiotic strains were sensitive to ampicillin, tetracycline, vancomycin and carbenicillin whereas most of the strains were resistant to erythromycin and novobiocin. Four of the 6 probiotic derived CFS possessed antimicrobial activity against C. sakazakii at a level of 40 μL. A higher biofilm inhibitory activity (>80%) was observed at initial stages of biofilm formation with weaker activity during longer incubation upto 48 hours (50%-60%).
CONCLUSIONS
The study indicated the efficacy of isolated commercial probiotics strains as potential inhibitor of biofilm formation by C. sakazakii and could be further explored for novel bioactive molecules to limit the emerging infections of C. sakazakii.

Keyword

Probiotics; Cronobacter sakazakii; Biofilm; Antimicrobial

MeSH Terms

Ampicillin
Biofilms*
Carbenicillin
Cronobacter sakazakii*
Cronobacter*
Drug Resistance, Microbial
Erythromycin
Gentian Violet
Humans
Infant
Infant, Newborn
Milk
Novobiocin
Probiotics*
Tetracycline
Vancomycin
Ampicillin
Carbenicillin
Erythromycin
Gentian Violet
Novobiocin
Tetracycline
Vancomycin

Figure

• 

  Fig. 1. (A-D) Effect of cell-free supernatant from probiotic cultures on biofilm formation by Cronobacter sakazakii stained with crystal violet staining (arrows indicate disrupted biofilm at different hours of incubation) at ×40.

• 

  Fig. 2. Fluorescent microscopic images of effect of cell-free supernatant (CFS) from probiotics on biofilms of Cronobacter sakazakii grown at different stages using live-dead stain at 50 μm. (A) Stained with SYTO-9, (B) slides stained with propidium Iodide (PI), (C) merged image.

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