Bioassay-guided isolation and identification of antibacterial compounds from Piper betle leaf with inhibitory activity against the Vibrio species in shrimp

ABSTRACT

The present study examined the antimicrobial effects of betel leaf (Piper betle) extracts against Vibrio spp. along with the isolation of the most prevalent bioactive components using a bioassay-guided purification. Following a preliminary screening, the ethanolic extract proved the most effective and performed to silica gel chromatography, followed by reversed phase HPLC to isolate the active components. By means of subsequent UHPLC-orbitrap-ion trap mass spectrometry, two compounds were identified as (E)-3-phenylprop-2-enoic acid (cinnamic acid) and [4-(1-hydroxyprop-2-enyl)phenyl] acetate (1’−hydroxychavicol acetate). The compounds showed anti-microbial activity against Vibrio species including V. parahaemolyticus, V. harveyi, V. alginolyticus, V. vulnificus and V. anguillarum. Their antibacterial action was quantified by a microdilution assay showing that cinnamic acid displays more potent activity (IC_50, 21–121 µg/mL) compared to 1’-hydroxychavicol acetate (IC_50, 24–365 µg/mL). These findings propose the potential use of the betel leaf as a substitute for antibiotics to combat Vibrio infections in shrimp aquaculture.

KEYWORDS:

• Piper betle
• bioassay-guided purification
• antimicrobial activity
• cinnamic acid
• 1’-hydroxychavicol acetate
• Vibrio

Acknowledgments

We are grateful to the Bangladesh National Herbarium (BNH), Government of the People’s Republic of Bangladesh, for assistance in plant identification. Sincere gratitude is extended to Professor Sebastien Carpentier and to Kusay Arat for assistance in using of their UHPLC-Orbitrap-Ion Trap Mass Spectrometer.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/10412905.2024.2353648

Credit author statement

Alokesh Kumar Ghosh: Conceptualization, Methodology, Software, Data curation, Writing-original draft preparation. Sujogya Kumar Panda: Supervision, Writing – review & editing. Haibo Hu: Software, Writing – review & editing. Liliane Schoofs: Supervision, Writing – review & editing, Walter Luyten: Supervision, Funding acquisition, Writing – review & editing.

Correction Statement

This article has been republished with minor changes. These changes do not impact the academic content of the article.

Additional information

Funding

First author A.K. Ghosh was funded by a Prime Minister Fellowship from the Government of the People’s Republic of Bangladesh. W. Luyten largely supported himself.