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1. Enhancing Antibacterial Efficacy: Synergistic Effects of Citrus aurantium Essential Oil Mixtures against Escherichia coli for Food Preservation

Author

Ines Ellouze, Boutheina Ben Akacha, Ivana Generalić Mekinić, Rania Ben Saad, Miroslava Kačániová, Maciej Ireneusz Kluz, Wissem Mnif, Stefania Garzoli, and Anis Ben Hsouna

Subjects
bitter orange, essential oils, chemical composition, synergistic effects, ternary mixture, Escherichia coli, Chemical technology, TP1-1185
Abstract

Essential oils (EOs) from various medicinal and aromatic plants are known for their diverse biological activities, including their antimicrobial effects. Citrus aurantium EO is traditionally used for therapeutic benefits due to its high content of bioactive compounds. Therefore, this study focuses on its potential use as a food preservative by investigating the combined antibacterial properties of EOs from leaves (EO1), flowers (EO2), and small branches (EO3) of Citrus aurantium against six bacterial strains by the agar disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) methods. The chemical compositions of the EOs were analysed by gas chromatography–mass spectrometry (GC-MS) and revealed the presence of numerous compounds responsible for their antimicrobial properties. The MIC values for the EOs were 3.12 mg/mL, 4.23 mg/mL, and 1.89 mg/mL, for EO1, EO2 and EO3, respectively, while the MBC values were 12.5 mg/mL, 6.25 mg/mL, and 6.25 mg/mL, respectively. A simplex centroid design was created to analyse the effect of the individual and combined EOs against E. coli. The combined EOs showed enhanced antibacterial activity compared to the individual oils, suggesting a synergistic effect (e.g., trial 9 with an MIC of 0.21 mg/mL), allowing the use of lower EO concentrations and reducing potential negative effects on food flavour and aroma. Additionally, the practical application of investigated EOs (at concentrations twice the MIC) was investigated in raw chicken meat stored at 4 °C for 21 days. The EOs, individually and in combination, effectively extended the shelf life of the meat by inhibiting bacterial growth (total bacterial count of less than 1 × 104 CFU/g in the treated samples compared to 7 × 107 CFU/g in the control on day 21 of storage). The study underlines the potential of C. aurantium EOs as natural preservatives that represent a sustainable and effective alternative to synthetic chemicals in food preservation.

Published
2024
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2. Season's variation impact on Citrus aurantium leaves essential oil: chemical composition and biological activities

Author

Nassereddine Sabaou, Florence Mathieu, Inès Ellouze, Manef Abderrabba, Jalloul Bouajila, Ahmed Lebrihi, Centre National de la Recherche Scientifique - CNRS (FRANCE), Ecole normale supérieure de Kouba (ALGERIA), Institut National Polytechnique de Toulouse - Toulouse INP (FRANCE), Université Toulouse III - Paul Sabatier - UT3 (FRANCE), Université de Carthage (TUNISIA), Laboratoire de Génie Chimique - LGC (Toulouse, France), Interactions moléculaires et réactivité chimique et photochimique (IMRCP), Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Fédération de Recherche Fluides, Energie, Réacteurs, Matériaux et Transferts (FERMAT), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université de Carthage - University of Carthage, École normale supérieure - Kouba-Alger (ENS Kouba-Alger), Laboratoire de Génie Chimique (LGC), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD), Laboratoire de génie chimique [ancien site de Basso-Cambo] (LGC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), and Université Fédérale Toulouse Midi-Pyrénées

Subjects
0106 biological sciences, Citrus, Biochimie, Biologie Moléculaire, Chemical composition, Linalyl acetate, 01 natural sciences, Antioxidants, Essential oil, law.invention, chemistry.chemical_compound, [CHIM.GENI]Chemical Sciences/Chemical engineering, Anti-Infective Agents, Fusarium, Linalool, law, Food science, Flame Ionization, 2. Zero hunger, 0303 health sciences, Antimicrobial, Biphenyl compound, Mucor, Seasons, Sesquiterpenes, Bacillus subtilis, Staphylococcus aureus, Acyclic Monoterpenes, Food Contamination, Cyclohexane Monoterpenes, Microbial Sensitivity Tests, Saccharomyces cerevisiae, Gas Chromatography-Mass Spectrometry, 03 medical and health sciences, Picrates, Cyclohexenes, Botany, Escherichia coli, Oils, Volatile, Génie chimique, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Phenols, Citrus aurantium, 030304 developmental biology, Polycyclic Sesquiterpenes, Biological activity, Biphenyl Compounds, Petitgrain, Plant Leaves, chemistry, 13. Climate action, Food Microbiology, Monoterpenes, CG/MS, Season, Gas chromatography–mass spectrometry, 010606 plant biology & botany, Food Science
Abstract

International audience; Citrus aurantium leaves' essential oils (EOs) were evaluated for chemical composition and antioxidant and antibacterial activities. The vegetable material, taken 5 times during the year, has undergone the hydrodistillation to prepare EO. Chemical characterization by gas chromatography/mass spectrometry and GC/flame ionization detection allowed the identification of 46 compounds, and a notable quantitative and qualitative differences between the different Petitgrain samples according to the harvest time. Linalool (43.2% to 65.97%), linalyl acetate (0.77% to 24.77%), and α-terpineol (9.29% to 12.12%) were the main components. The most important number of components was registered for summer EOs (July and September). The 5 EOs submitted biological activities screening, namely, antioxidant and antimicrobial activities. Weak antioxidant activities (IC50 values >10000 mg/L) were registered by both 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis-3-ethylbenzothiazoline-6-sulphonate assays, mostly because the weak amount of phenols in EOs. Antibacterial activities (12 microorganisms) were registered against Gram-positive bacteria [Bacillus subtilis (MIC = 2.7 mg/mL), Staphylococcus aureus (4.8 mg/mL)], and moderated ones against yeasts [Saccharomyces cerevisiae (9.2 mg/mL)] and fungi [Mucor ramannianus (5 mg/mL)]. Positive correlations between the identified compounds and the antimicrobial activities were noted. Many compounds were correlated to antimicrobial activity mainly caryophyllene oxide against Escherichia coli (R² = 0.99), S. cerevisiae (R² = 0.99), and Fusarium culmorum (R² = 0.99).

Published
2012
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