Sub-inhibitory concentrations of gentamicin triggers the expression of aac(6′)Ie-aph(2″)Ia, chaperons and biofilm related genes in Lactobacillus plantarum MCC 3011

Jaimee, George; Halami, Prakash M.

doi:10.1016/j.resmic.2017.06.002

Cited by 14 publications

(5 citation statements)

References 37 publications

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“…However, most of the previous studies have been performed on pathogens (Jahn et al, 2017). L. plantarum could gain high level resistance after constant exposure to aminoglycosides (George and Halami, 2017), particularly L. plantarum had good adaptability to gentamicin and kanamycin with an increasing of antibiotic concentration during the passage. In this work, a probiotic L. plantarum strain was selected to perform a laboratory evolution study to better understand the impact of a gentamicin selective pressure on this bacterial species at whole-genome level.…”

Section: Discussionmentioning

confidence: 99%

Laboratory Evolution Assays and Whole-Genome Sequencing for the Development and Safety Evaluation of Lactobacillus plantarum With Stable Resistance to Gentamicin

et al. 2019

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The goal of this work was to use laboratory evolution assays and whole-genome sequencing to develop and test the safety of a probiotic, Lactobacillus plantarum , with high-level of resistance to gentamicin. The evolution of L. plantarum was evaluated under the selective pressure from gentamicin and subsequently when the selective pressure was removed. After 30 days of selective pressure from gentamicin, the minimum inhibitory concentration (MIC) of L. plantarum to gentamicin increased from 4 to 512 μg/mL and remained stable at this level. After removing the selective pressure, the resistance of L. plantarum to gentamicin decreased to 64 μg/mL after 20 days, and remained stable thereafter. Although the MIC declined it was still higher than the cut-off value recommended by EFSA, indicating that the acquisition of gentamicin-resistance was an irreversible process. Using whole-genome sequencing, gene mutations were identified in the strains that had undergone selection pressure from gentamicin as well as in the strains where the selection pressure was subsequently removed. Specifically, four non-synonymous mutations were detected including one single nucleotide polymorphism (SNP), one insertion, and two structural variants (SVs), of which the mutations in genes encoding the drug resistance MFS transporter and transcriptional regulator of AraC family were only detected in the strains under selective pressure from gentamicin. The results indicate that these mutations play an important role in increasing the resistant levels of L. plantarum to gentamicin. The mobility analysis of mutant genes confirmed that they were not located on mobile elements of the genome of highly resistant L. plantarum , indicating that horizontal gene transfer was not possible.

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Section: Discussionmentioning

confidence: 99%

Laboratory Evolution Assays and Whole-Genome Sequencing for the Development and Safety Evaluation of Lactobacillus plantarum With Stable Resistance to Gentamicin

et al. 2019

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“…Residual concentrations of excreted gentamicin have been detected in wastewater treatment plants [ 21 ]. Gentamicin at sub-inhibitory concentrations has been shown to induce resistance development in pure cultures [ 22 ]. Moreover, aminoglycoside resistance genes are widely distributed in both chromosomal and plasmid class 1 integrons [ 23 ], which are considered major vectors of antibiotic resistance dissemination [ 24 ].…”

Section: Introductionmentioning

confidence: 99%

Gentamicin at sub-inhibitory concentrations selects for antibiotic resistance in the environment

Sanchez-Cid

Guironnet

Keuschnig

et al. 2022

ISME Communications

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Antibiotics released into the environment at low (sub-inhibitory) concentrations could select for antibiotic resistance that might disseminate to the human microbiome. In this case, low-level anthropogenic sources of antibiotics would have a significant impact on human health risk. In order to provide data necessary for the evaluation of this risk, we implemented river water microcosms at both sub-inhibitory and inhibitory concentrations of gentamicin as determined previously based on bacterial growth in enriched media. Using metagenomic sequencing and qPCR/RT-qPCR, we assessed the effects of gentamicin on water bacterial communities and their resistome. A change in the composition of total and active communities, as well as a gentamicin resistance gene selection identified via mobile genetic elements, was observed during a two-day exposure. We demonstrated the effects of sub-inhibitory concentrations of gentamicin on bacterial communities and their associated resistome in microcosms (simulating in situ conditions). In addition, we established relationships between antibiotic dose and the magnitude of the community response in the environment. The scope of resistance selection under sub-inhibitory concentrations of antibiotics and the mechanisms underlying this process might provide the basis for understanding resistance dispersion and associated risks in relatively low impacted ecosystems.

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“…L. plantarum MCC2774, L. plantarum CSG-8, L. plantarum CSG-21, L. plantarum C26a and L. plantarum MCC3011 isolated from chicken sausages were subjected to minimum inhibitory concentration (MIC) determination28. These were also found to possess the bi-functional aminoglycoside resistance gene15. The native cultures were grown and maintained in LAB susceptibility medium (LSM) and were used in this study.…”

Section: Methodsmentioning

confidence: 99%

“…Biofilms have been proposed to play a major role in deciphering antibiotic resistance, especially aminoglycosides15. The existence of a close microbial niche within biofilms and varying genetic responses are caused due to the density-dependent mechanism.…”

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confidence: 99%

Presence of extracellular DNA & protein in biofilm formation by gentamicin-resistant Lactobacillus plantarum

Jaimee

Halami

2019

Indian Journal of Medical Research

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Background & objectives: Bacterial biofilms a multi-layered defence, comprise extracellular DNA (eDNA) and proteins, protect bacteria from harmful environment and nutrient limitation and utilize the mutual benefits within a community. Bacterial biofilms also defend bacteria from harsh environments such as antibiotic treatment. This leads to poor antibiotic penetration, slow growth, adaptive stress responses, and formation of persister cells. This study was done to determine the relation of antibiotic resistance deciphered by the biofilms in Lactobacillus plantarum , a lactic acid bacteria (LAB) with probiotic significance. Methods: The gentamicin-resistant L. plantarum isolates were allowed to form biofilms and subjected to DNase I and proteinase K treatment. The optical density (OD) values were recorded for the biofilm assay and the cell count for the number of viable cells was taken for the control and the test samples. Percentage reduction was calculated based on the difference between the initial and final OD for both the parameters. Results: The biofilm assay revealed that the native L. plantarum isolates which were phenotypically susceptible, possessed the ability to form biofilms. The OD values were significantly decreased in comparison to the biofilm-forming control culture when these were treated with DNase I and proteinase K. Interpretation & conclusions: The study revealed that the biofilms formed by L. plantarum comprised of eDNA and proteins which was evidenced by the reduction in OD values and percentage in comparison to the control upon DNase I and proteinase K treatment. This indicates that the eDNA and biofilm matrix proteins are vital constituents of biofilms and may carry significant risk when coupled with antibiotic resistance.

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Sub-inhibitory concentrations of gentamicin triggers the expression of aac(6′)Ie-aph(2″)Ia, chaperons and biofilm related genes in Lactobacillus plantarum MCC 3011

Cited by 14 publications

References 37 publications

Laboratory Evolution Assays and Whole-Genome Sequencing for the Development and Safety Evaluation of Lactobacillus plantarum With Stable Resistance to Gentamicin

Laboratory Evolution Assays and Whole-Genome Sequencing for the Development and Safety Evaluation of Lactobacillus plantarum With Stable Resistance to Gentamicin

Gentamicin at sub-inhibitory concentrations selects for antibiotic resistance in the environment

Presence of extracellular DNA & protein in biofilm formation by gentamicin-resistant Lactobacillus plantarum

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