Plasmid stability of potential probiotic Lactobacillus plantarum strains in artificial gastric juice, at elevated temperature, and in the presence of novobiocin and acriflavine

Arch Microbiol. 2021 Jan;203(1):183-191. doi: 10.1007/s00203-020-02017-4. Epub 2020 Aug 16.


In this study, the presence of plasmids responsible for carbohydrate fermentation and antibiotic resistance and the stability of these plasmids in artificial gastric juice were investigated in 20 Lactobacillus plantarum strains with probiotic properties. Plasmid curing was performed with novobiocin, acriflavine and elevated incubation temperature to identify plasmids encoded with carbohydrate fermentation and antibiotic resistance genes and to compare them with artificial gastric juice. Plasmid profiling of the strains revealed that 100% of the strains were harbouring plasmids in varying sizes and numbers. The plasmid number of the potential probiotic strains ranged between 1 and 4, and the plasmid size ranged between 5.779 and 16.138 kb. The potential probiotic strains could not survive in the artificial gastric juice at pH 2.0. Although the strains maintained their viability in an artificial gastric juice at pH 2.5 and 3.0, and their derivatives lost their plasmids at a high rate (100%). Similarly, high levels of cured derivatives were obtained with 8 µg/mL novobiocin and 100 µg/mL acriflavine applications, and 24 h incubation at 43 °C. All the experiments were also performed to compare with two L. plantarum-type strains containing plasmids responsible for tetracycline and tetracycline + erythromycin resistances. Artificial gastric juice and other plasmid curing treatments caused a high-frequency loss in the antibiotic resistances of type strains. Determining plasmid stability in artificial gastric juice is a novel approach. Plasmid stability in the gastrointestinal tract is important for maintaining the plasmid-encoded probiotic properties.

Keywords: Antibiotic resistances; Artificial gastric juice; Carbohydrate fermentation; Plasmid stability; Probiotics.

MeSH terms

  • Acriflavine / pharmacology*
  • Anti-Bacterial Agents / pharmacology
  • Drug Resistance, Bacterial / genetics*
  • Fermentation
  • Gastric Juice / drug effects
  • Gastric Juice / microbiology*
  • Hot Temperature
  • Lactobacillus plantarum / drug effects*
  • Lactobacillus plantarum / genetics
  • Novobiocin / pharmacology*
  • Plasmids / genetics
  • Probiotics
  • Tetracycline Resistance / genetics


  • Anti-Bacterial Agents
  • Novobiocin
  • Acriflavine