Antibiotic resistance modulation and modes of action of (-)-α-Pinene in Campylobacter jejuni

Jasna Kovač, Katarina Šimunović, Zuowei Wu, Anja Klančnik, Franz Bucar, Qijing Zhang, Sonja Smole Možina

Research output: Contribution to journalArticle

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Abstract

The aim of the study was to investigate the mode of action of (-)-α-pinene in terms of its modulation of antibiotic resistance in Campylobacter jejuni. Broth microdilution and ethidium bromide accumulation assays were used to evaluate the (-)-α -pinene antimicrobial activity, modulation of antimicrobial resistance, and inhibition of antimicrobial efflux. The target antimicrobial efflux systems were identified using an insertion mutagenesis approach, and C. jejuni adaptation to (-)-α-pinene was evaluated using DNA microarrays. Knock-out mutants of the key up-regulated transcriptional regulators hspR and hrcA were constructed to investigate their roles in C. jejuni adaptation to several stress factors, including osmolytes, and pH, using Biolog phenotypical microarrays. Our data demonstrate that (-)- α-pinene efficiently modulates antibiotic resistance in C. jejuni by decreasing the minimum inhibitory concentrations of ciprofloxacin, erythromycin and triclosan by up to 512-fold. Furthermore, (-)- α-pinene promotes increased expression of cmeABC and another putative antimicrobial efflux gene, Cj1687. The ethidium bromide accumulation was greater in the wild-type strain than in the antimicrobial efflux mutant strains, which indicates that these antimicrobial efflux systems are a target of action of (-)- α-pinene. Additionally, (-)-α-pinene decreases membrane integrity, which suggests that enhanced microbial influx is a secondary mode of action of (-)-α-pinene. Transcriptomic analysis indicated that (-)-α-pinene disrupts multiple metabolic pathways, and particularly those involved in heat-shock responses. Thus, (-)- α-pinene has significant activity in the modulation of antibiotic resistance in C. jejuni, which appears to be mediated by multiple mechanisms that include inhibition of microbial efflux, decreased membrane integrity, and metabolic disruption. These data warrant further studies on (-)-α-pinene to develop its use in the control of antibiotic resistance in Campylobacter.

Original languageEnglish (US)
Article numbere0122871
JournalPloS one
Volume10
Issue number4
DOIs
StatePublished - Apr 1 2015

Fingerprint

Campylobacter jejuni
Microbial Drug Resistance
antibiotic resistance
mechanism of action
anti-infective agents
Modulation
Anti-Bacterial Agents
Ethidium
Microarrays
Triclosan
Membranes
Mutagenesis
Erythromycin
Ciprofloxacin
Heat-Shock Response
Campylobacter
Microbial Sensitivity Tests
Assays
Metabolic Networks and Pathways
Oligonucleotide Array Sequence Analysis

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Kovač, J., Šimunović, K., Wu, Z., Klančnik, A., Bucar, F., Zhang, Q., & Možina, S. S. (2015). Antibiotic resistance modulation and modes of action of (-)-α-Pinene in Campylobacter jejuni. PloS one, 10(4), [e0122871]. https://doi.org/10.1371/journal.pone.0122871
Kovač, Jasna ; Šimunović, Katarina ; Wu, Zuowei ; Klančnik, Anja ; Bucar, Franz ; Zhang, Qijing ; Možina, Sonja Smole. / Antibiotic resistance modulation and modes of action of (-)-α-Pinene in Campylobacter jejuni. In: PloS one. 2015 ; Vol. 10, No. 4.
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Kovač, J, Šimunović, K, Wu, Z, Klančnik, A, Bucar, F, Zhang, Q & Možina, SS 2015, 'Antibiotic resistance modulation and modes of action of (-)-α-Pinene in Campylobacter jejuni', PloS one, vol. 10, no. 4, e0122871. https://doi.org/10.1371/journal.pone.0122871

Antibiotic resistance modulation and modes of action of (-)-α-Pinene in Campylobacter jejuni. / Kovač, Jasna; Šimunović, Katarina; Wu, Zuowei; Klančnik, Anja; Bucar, Franz; Zhang, Qijing; Možina, Sonja Smole.

In: PloS one, Vol. 10, No. 4, e0122871, 01.04.2015.

Research output: Contribution to journalArticle

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