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ARS Home » Southeast Area » Fayetteville, Arkansas » Poultry Production and Product Safety Research » Research » Publications at this Location » Publication #339666

Research Project: Antibiotic Alternatives for Controlling Foodborne Pathogens and Disease in Poultry

Location: Poultry Production and Product Safety Research

Title: Plant-derived antimicrobial eugenol modulates C. jejuni proteome and virulence critical for colonization in chickens

item UPADHYAYA, INDU - University Of Arkansas
item UPADHYAY, ABHINAV - University Of Arkansas
item ARSI, KOMALA - University Of Arkansas
item LIYANAGE, ROHANA - University Of Arkansas
item Donoghue, Ann - Annie
item Rath, Narayan
item DONOGHUE, DAN - University Of Arkansas

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 5/2/2017
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Campylobacter jejuni is an important foodborne pathogen that causes severe diarrhea in humans. Chickens act as the reservoir host for Campylobacter, wherein the pathogen colonizes the ceca leading to contaminated poultry products during slaughter. The potential of natural intervention strategies, including plant-derived antimicrobials have been investigated to reduce chicken cecal colonization of C. jejuni. However, the underlying molecular mechanisms of these strategies are still unknown. This study investigated the effect of eugenol (a generally recognized as safe status phytochemical obtained from clove oil), on the whole cell proteomic profile of C. jejuni. In addition, the effect of eugenol on factors critical for cecal colonization (bacterial motility, attachment to epithelial cells) in chickens were studied using 0.4% agar for a motility assay and cell culture analysis using chicken primary enterocytes isolated from broilers. C. jejuni (strain S-8, isolated from commercial broilers) was cultured in the presence or absence (control) of sub-inhibitory concentrations of eugenol (0.01%) for 24 h followed by SDS-PAGE based protein extraction. The effect of eugenol on expression of C. jejuni proteome was quantified using LC-MS/MS analysis followed by targeted proteomics and data analysis using Scaffold Proteomic software. The experiment was conducted three times and the samples were run in triplicates. Whole cell proteomic analysis identified more than 600 proteins in C. jejuni with many virulent proteins modulated by eugenol. The major groups of proteins that were identified contribute to physiological process (sensory systems, biological regulators, developmental processes) and virulent attributes (motility systems, adhesion, quorum sensing) in C. jejuni. Eugenol reduced the expression of major virulence proteins contributing to biological adhesion (PorA, CadF), motility system (MotA, MotB, FliA, FliD, FliF, fliL, fliY), energy taxis (IlvH, CetA, CetB), molecular transport (TatA, TatB, TolB) and Quorum sensing (LuxS) when compared to controls (P<0.05). Follow up motility and attachment assays revealed that eugenol was effective in reducing the motility (> 50% reduction) and attachment of C. jejuni to primary chicken enterocytes (P<0.05). Overall, these results delineate the prospective mechanism of action of eugenol on C. jejuni and the potential of using this phytochemical to control C. jejuni colonization in chickens.