Development of Resistance to Clarithromycin and Amoxicillin-Clavulanic Acid in Lactiplantibacillus plantarum In Vitro Is Followed by Genomic Rearrangements and Evolution of Virulence

Abstract

Ensuring the safety of the use of probiotics is a top priority. We found that increased resistance to popular antimicrobial drugs in Lactiplantibacillus plantarum is accompanied by significant changes in the genomic profile and phenotypic sensitivity to a number of antimicrobial drugs as well as in the level of virulence of this bacterium against Drosophila .

Repression of Staphylococcus aureus and Escherichia coli by Lactiplantibacillus plantarum Strain AG10 in Drosophila melanogaster In Vivo Model

“…Since bacteria added to the flies’ growth media were preliminary washed by PBS, the observed effects are apparently driven by antimicrobial substances produced by Lactobacilli de novo. Moreover, the experiment with S. aureus and E. coli overexpressing GFP confirmed that pathogens are captured by larvae ( Figure 2 ), whereas successful introduction of Lactobacilli to the larvaes’ gut was shown previously [ 100 ], suggesting that the repression of pathogens occurs in vivo, while ex vivo interactions remain also possible.…”

“…Further analysis of the L. plantarum AG10 genome using the VFDB database revealed, in silico, only three putative virulence factors ( Table 6 ), while 30 were predicted for L. plantarum 8PA3 [ 100 ]; this apparently suggests the safety of L. plantarum AG10 as a probiotic strain.…”

“…Similar suppression has been observed for S. aureus under the same conditions (Compare Figure 1 C,D), suggesting antimicrobial potential of the strain. In addition, low amounts of putative virulence factors have been detected in silico in the genome of L. plantarum AG10 by using the VFDB database (in contrast to 30 found in L. plantarum 8PA3 [ 100 ]), suggesting its prominent safety profile as a potential probiotic.…”

Accounting for the health risk of probiotics

“…Since BKGI1 homologs are also present in other bifidobacterial strains especially B. longum , the genomic island BKGI1 therefore mediates the spread and integration of erm(X) in bifidobacteria [ 185 ]. Adaptive evolution of Lactiplantibacillus plantarum isolated from a commercially available probiotic health product led to a daughter strain with increased resistance to amoxicillin-clavulanic acid and clarithromycin which are used to treat Helicobacter pylori infections [ 186 ]. The increased resistance to clarithromycin and amoxicillin-clavulanic acid is also accompanied by the development of resistance to other classes of antibiotics such as fluoroquinolones and cephalosporins but resulted in a decrease in resistance to aminoglycosides, tetracyclines, and rifampicin.…”

“…Comparative genomic analysis of the adapted antibiotic-resistant and parental strains detected point mutations and larger-scale genomic rearrangements that may account for the development of resistance such as the insertion of the transposase gene ISLpL3 into the esterase gene. Interestingly, the antibiotic-resistance strain is accompanied by an increased in virulence in Drosophila melanogaster which is a natural host for the L. plantarum symbiont, assessed through viability and reproduction studies such as egg numbers, embryonic death, and DNA damage in enterocytes of the flies [ 186 ].…”