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In vitro and in vivo characterization of citral and limonene combinations against Vibrio campbellii in brine shrimp
Zheng, X.; Declercq, A.M.; Vanrompay, D.; Feyaerts, A.F.; Van Dijck, P.; Bossier, P. (2024). In vitro and in vivo characterization of citral and limonene combinations against Vibrio campbellii in brine shrimp. Aquaculture 593: 741343. https://dx.doi.org/10.1016/j.aquaculture.2024.741343
In: Aquaculture. Elsevier: Amsterdam; London; New York; Oxford; Tokyo. ISSN 0044-8486; e-ISSN 1873-5622, more
Peer reviewed article  

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Keywords
    Vibrio campbellii (Baumann, Baumann & Mandel, 1971) Baumann, Baumann, Bang & Woolkalis, 1981 [WoRMS]
    Marine/Coastal
Author keywords
    Citral; Limonene; Brine shrimp; Essential oil component

Authors  Top 
  • Feyaerts, A.F.
  • Van Dijck, P., more
  • Bossier, P., more

Abstract
    The use of essential oils (EOs) has recently gained popularity due to their antimicrobial properties. However, current research primarily focuses on basic laboratory tests to identify active compounds or screen different plant sources. EOs comprise numerous constituents that interact intricately, complicating the understanding of their effective mechanisms. To address this issue, the study aimed to investigate the antibacterial activity and biological function of two major components (citral and limonene) extracted from the EO of Litsea citrata. Six assays were performed to investigate the antibacterial activity of citral and limonene, including a checkerboard assay, an inhibitory growth assay, a vapor-phase-mediated assay, a regrowth assay, a release of cellular contents assay, and an in vivo bioluminescence assay. Additionally, a challenge test and quantitative PCR of immune gene expression were conducted to assess the biological function of citral and limonene. The results showed that a citral/limonene mixture in a 1:8 ratio exhibited a synergistic effect against V. campbellii. While the antibacterial activity of the EOC mixture of citral/limonene (1:8) at the concentration of 0.0005% was relatively weak, it significantly increased the survival of Artemia when challenged with V. campbellii in vivo. These results suggest that the citral/limonene mixture in a 1:8 ratio at 0.0005% probably protects brine shrimp by immunomodulation at the host level and by reducing the colonization and/or virulence of the pathogen V. campbellii. Our results indicate the potential of this novel EOC combination to combat vibriosis in aquaculture.

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