(-)-α-Pinene reduces quorum sensing and Campylobacter jejuni colonization in broiler chickens

PLoS One. 2020 Apr 1;15(4):e0230423. doi: 10.1371/journal.pone.0230423. eCollection 2020.

Abstract

Campylobacter jejuni is one of the most prevalent causes of bacterial gastroenteritis worldwide, and it is largely associated with consumption of contaminated poultry. Current Campylobacter control measures at the poultry production level remain insufficient, and hence there is the need for alternative control strategies. We evaluated the potential of the monoterpene (-)-α-pinene for control of C. jejuni in poultry. The antibacterial and resistance-modulatory activities of (-)-α-pinene were also determined against 57 C. jejuni strains. In addition, the anti-quorum-sensing activity of (-)-α-pinene against C. jejuni NCTC 11168 was determined for three subinhibitory concentrations (125, 62.5, 31.25 mg/L) over three incubation times using an autoinducer-2 bioassay based on Vibrio harveyi BB170 bioluminescence measurements. The effects of a subinhibitory concentration of (-)-α-pinene (250 mg/L) on survival of C. jejuni, and in combination with enrofloxacin on fluoroquinolone resistance development in C. jejuni, were determined in a broiler chicken model, by addition of (-)-α-pinene to the broiler water supply. The reduction of C. jejuni numbers by (-)-α-pinene was further determined in broiler chickens that were colonized with either fluoroquinolone-susceptible or -resistant strains, by direct gavage treatment. We observed weak in vitro antimicrobial activity for (-)-α-pinene alone (MIC >500 mg/L), but strong potentiating effects on antibiotics erythromycin and ciprofloxacin against different Campylobacter strains (>512 fold change). After 24 h of treatment of C. jejuni with (-)-α-pinene, its quorum-sensing signaling was reduced by >80% compared to the untreated control. When given in the drinking water, (-)-α-pinene did not show any significant inhibitory effects on the level of C. jejuni in the colonized chickens, and did not reduce fluoroquinolone resistance development in combination with enrofloxacin. Conversely, when (-)-α-pinene was administered by direct gavage, it significantly reduced the number of fluoroquinolone susceptible C. jejuni in the colonized broiler chickens. These results demonstrate that (-)-α-pinene modulates quorum-sensing in Campylobacter, potentiates antibiotics against different Campylobacter strains, and reduces Campylobacter colonization in broiler chickens.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Anti-Bacterial Agents / pharmacology*
  • Anti-Bacterial Agents / therapeutic use
  • Bicyclic Monoterpenes / pharmacology*
  • Bicyclic Monoterpenes / therapeutic use
  • Campylobacter Infections / pathology
  • Campylobacter Infections / prevention & control
  • Campylobacter jejuni / drug effects*
  • Campylobacter jejuni / physiology
  • Cecum / microbiology
  • Chickens
  • Ciprofloxacin / pharmacology
  • Drug Resistance, Bacterial / drug effects
  • Erythromycin / pharmacology
  • Fluoroquinolones / pharmacology
  • Fluoroquinolones / therapeutic use
  • Microbial Sensitivity Tests
  • Poultry Diseases / pathology
  • Poultry Diseases / prevention & control
  • Quorum Sensing / drug effects*

Substances

  • Anti-Bacterial Agents
  • Bicyclic Monoterpenes
  • Fluoroquinolones
  • Ciprofloxacin
  • Erythromycin
  • alpha-pinene

Associated data

  • figshare/10.6084/m9.figshare.8234153
  • figshare/10.6084/m9.figshare.8234192
  • figshare/10.6084/m9.figshare.8234189
  • figshare/10.6084/m9.figshare.8234156
  • figshare/10.6084/m9.figshare.8234159

Grants and funding

This study was financed by the Slovenian-American bilateral projects BI-SLO-USA 2014/2015 and 2018/19 and P4-0116 funded by the Slovenian Research Agency (ARRS) awarded to SSM. http://www.arrs.si/en/index.asp JK was supported by the USDA National Institute of Food and Hatch Appropriations under Project #PEN04646 and Accession #1015787. https://nifa.usda.gov/ The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.