Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step Development
Essential oils of <i>Citrus sinensis</i> and <i>Citrus latifolia</i> have shown biological functions as antiseptics, anti-inflammatories, antioxidants, antifungal and antimutagenic, so the evaluation of their antibacterial capacity, by themselves or in combination with standa...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-10-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/10/10/1218 |
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| author | Nayeli G. Nieto-Velázquez Alejandra A. Gomez-Valdez Marisela González-Ávila Jaime Sánchez-Navarrete Julia D. Toscano-Garibay Nancy J. Ruiz-Pérez |
| author_facet | Nayeli G. Nieto-Velázquez Alejandra A. Gomez-Valdez Marisela González-Ávila Jaime Sánchez-Navarrete Julia D. Toscano-Garibay Nancy J. Ruiz-Pérez |
| author_sort | Nayeli G. Nieto-Velázquez |
| collection | DOAJ |
| description | Essential oils of <i>Citrus sinensis</i> and <i>Citrus latifolia</i> have shown biological functions as antiseptics, anti-inflammatories, antioxidants, antifungal and antimutagenic, so the evaluation of their antibacterial capacity, by themselves or in combination with standard antibiotics, presents an alternative for infection treatment. Flow cytometry opens the door for the design of faster and more accurate measurement of antibacterial activity. We use a SYTO9/PI staining system on <i>E. coli</i> ATCC 25922 to determine antibacterial activity by counting live and dead cells through flow cytometry. We found that dual staining showed highly variable results due to wavelength overlapping and instead we used fluorochrome individual staining that highly correlated with viable counts. Chloramphenicol and cefotaxime treatments did not present a dose-response behavior, rendered diffuse readings and/or gave filament formation on fluorescence microscopy. Amikacin was a better comparison standard because it presented a dose-response behavior. Essential oils had low antibacterial activity as compared to amikacin, with a maximum of 10% and 20% for <i>C. latifolia</i> and <i>C. sinensis</i>, respectively. Combinations of essential oils with antibiotic resulted in an unforeseen strong inhibition of amikacin activity. Although a low antibacterial activity was found, a series of standardization steps are proposed for antibacterial activity measurement by flow cytometry. |
| first_indexed | 2024-03-10T06:46:11Z |
| format | Article |
| id | doaj.art-53b7954f68c94907aba645aaac4194be |
| institution | Directory Open Access Journal |
| issn | 2079-6382 |
| language | English |
| last_indexed | 2024-03-10T06:46:11Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj.art-53b7954f68c94907aba645aaac4194be2023-11-22T17:13:56ZengMDPI AGAntibiotics2079-63822021-10-011010121810.3390/antibiotics10101218Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step DevelopmentNayeli G. Nieto-Velázquez0Alejandra A. Gomez-Valdez1Marisela González-Ávila2Jaime Sánchez-Navarrete3Julia D. Toscano-Garibay4Nancy J. Ruiz-Pérez5Dirección de Investigación y Enseñanza, Hospital Juárez de México, Ciudad de México 07760, MexicoFacultad de Estudios Superiores Cuatitlán, Universidad Nacional Autónoma de México, Cuautitlán Izcalli 54740, MexicoCentro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Guadalajara 44270, MexicoUnidad de Banco de Sangre, Hospital Juárez de México, Ciudad de México 07760, MexicoDirección de Investigación y Enseñanza, Hospital Juárez de México, Ciudad de México 07760, MexicoDirección de Investigación y Enseñanza, Hospital Juárez de México, Ciudad de México 07760, MexicoEssential oils of <i>Citrus sinensis</i> and <i>Citrus latifolia</i> have shown biological functions as antiseptics, anti-inflammatories, antioxidants, antifungal and antimutagenic, so the evaluation of their antibacterial capacity, by themselves or in combination with standard antibiotics, presents an alternative for infection treatment. Flow cytometry opens the door for the design of faster and more accurate measurement of antibacterial activity. We use a SYTO9/PI staining system on <i>E. coli</i> ATCC 25922 to determine antibacterial activity by counting live and dead cells through flow cytometry. We found that dual staining showed highly variable results due to wavelength overlapping and instead we used fluorochrome individual staining that highly correlated with viable counts. Chloramphenicol and cefotaxime treatments did not present a dose-response behavior, rendered diffuse readings and/or gave filament formation on fluorescence microscopy. Amikacin was a better comparison standard because it presented a dose-response behavior. Essential oils had low antibacterial activity as compared to amikacin, with a maximum of 10% and 20% for <i>C. latifolia</i> and <i>C. sinensis</i>, respectively. Combinations of essential oils with antibiotic resulted in an unforeseen strong inhibition of amikacin activity. Although a low antibacterial activity was found, a series of standardization steps are proposed for antibacterial activity measurement by flow cytometry.https://www.mdpi.com/2079-6382/10/10/1218resistance<i>E. coli</i>flow cytometryessential oilscitrusviability |
| spellingShingle | Nayeli G. Nieto-Velázquez Alejandra A. Gomez-Valdez Marisela González-Ávila Jaime Sánchez-Navarrete Julia D. Toscano-Garibay Nancy J. Ruiz-Pérez Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step Development Antibiotics resistance <i>E. coli</i> flow cytometry essential oils citrus viability |
| title | Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step Development |
| title_full | Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step Development |
| title_fullStr | Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step Development |
| title_full_unstemmed | Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step Development |
| title_short | Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step Development |
| title_sort | preliminary study on citrus oils antibacterial activity measured by flow cytometry a step by step development |
| topic | resistance <i>E. coli</i> flow cytometry essential oils citrus viability |
| url | https://www.mdpi.com/2079-6382/10/10/1218 |
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