Piper betle and its bioactive metabolite phytol mitigates quorum sensing mediated virulence factors and biofilm of nosocomial pathogen Serratia marcescens in vitro

Piper betle, a tropical creeper plant belongs to the family Piperaceae. The leaves of this plant have been well known for their therapeutic, religious and ceremonial value in South and Southeast Asia. It has also been reported to possess several biological activities including antimicrobial, antioxi...

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Published in:Journal of ethnopharmacology 2016-12, Vol.193, p.592-603
Main Authors: Srinivasan, Ramanathan, Devi, Kannan Rama, Kannappan, Arunachalam, Pandian, Shunmugiah Karutha, Ravi, Arumugam Veera
Format: Article
Language:English
Subjects:
Biofilm
Biofilms - drug effects
Biofilms - growth & development
Biomass
Cross Infection - microbiology
Cross Infection - urine
Dose-Response Relationship, Drug
Humans
Microscopy, Confocal
Microscopy, Electron, Scanning
Nosocomial infections
Phytol
Phytol - isolation & purification
Phytol - pharmacology
Piper betle
Piper betle - chemistry
Piper betle - metabolism
Plant Leaves - chemistry
Plant Leaves - metabolism
Prodigiosin - antagonists & inhibitors
Quorum sensing
Quorum Sensing - drug effects
Serratia marcescens
Serratia marcescens - drug effects
Serratia marcescens - growth & development
Serratia marcescens - metabolism
Serratia marcescens - pathogenicity
Virulence
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Summary:Piper betle, a tropical creeper plant belongs to the family Piperaceae. The leaves of this plant have been well known for their therapeutic, religious and ceremonial value in South and Southeast Asia. It has also been reported to possess several biological activities including antimicrobial, antioxidant, antinociceptive, antidiabetic, insecticidal and gastroprotective activities and used as a common ingredient in indigenous medicines. In Indian system of ayurvedic medicine, P. betle has been well recognized for its antiseptic properties and is commonly applied on wounds and lesions for its healing effects. To evaluate the anti-quorum sensing (anti-QS) and antibiofilm efficacy of P. betle and its bioactive metabolite phytol against Serratia marcescens. The P. betle ethyl acetate extract (PBE) was evaluated for its anti-QS efficacy against S. marcescens by assessing the prodigiosin and lipase production at 400 and 500µgml−1 concentrations. In addition, the biofilm biomass quantification assay was performed to evaluate the antibiofilm activity of PBE against S. marcescens. Besides, the influence of PBE on bacterial biofilm formation was assessed through microscopic techniques. The biofilm related phenomenons like exopolysaccharides (EPS) production, hydrophobicity and swarming motility were also examined to support the antibiofilm activity of PBE. Transcriptional analysis of QS regulated genes in S. marcescens was also done. Characterization of PBE was done by separation through column chromatography and identification of active metabolites by gas chromatography -mass spectrometry. The major compounds of active fractions such as hexadecanoic acid, eugenol and phytol were assessed for their anti-QS activity against S. marcescens. Further, the in vitro bioassays such as protease, biofilm and HI quantification were also carried out to confirm the anti-QS and antibiofilm potential of phytol in PBE. PBE inhibits QS mediated prodigiosin pigment production in S. marcescens, which confirmed its anti-QS potential against S. marcescens. At 500µgml−1 concentration, PBE significantly inhibited the production of protease, lipase, biofilm and EPS to the level of 71%, 68%, 65% and 43% in S. marcescens, respectively. Further, their antibiofilm efficacy was confirmed through microscopic techniques. In addition, PBE effectively inhibited the hydrophobicity and swarming motility. Additionally, the results of qPCR analysis validated the downregulation of QS genes. Chromatographi
ISSN:0378-8741
1872-7573
DOI:10.1016/j.jep.2016.10.017