Pheromone Activity after Stimulation with Ampicillin in a Plasmid-Free <i>Enterococcus faecalis</i> Strain
Enterococci exhibit clumping under the selective pressure of antibiotics. The aim of this study was to analyze the effect of supernatants from a plasmid-free clone (C29) of <i>Enterococcus faecalis</i> subjected to 0.25×, 0.5×, and 0.75× of the minimal inhibitory concentration (MIC) of a...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/10/11/2294 |
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| author | José Arellano-Galindo Sergio Zavala-Vega Rosario Vázquez-Larios Sara A. Ochoa Ariadnna Cruz-Córdova Adolfo Sierra-Santoyo Lourdes López-González Rigoberto Hernández-Castro Silvia Giono-Cerezo Juan Xicohtencatl-Cortes |
| author_facet | José Arellano-Galindo Sergio Zavala-Vega Rosario Vázquez-Larios Sara A. Ochoa Ariadnna Cruz-Córdova Adolfo Sierra-Santoyo Lourdes López-González Rigoberto Hernández-Castro Silvia Giono-Cerezo Juan Xicohtencatl-Cortes |
| author_sort | José Arellano-Galindo |
| collection | DOAJ |
| description | Enterococci exhibit clumping under the selective pressure of antibiotics. The aim of this study was to analyze the effect of supernatants from a plasmid-free clone (C29) of <i>Enterococcus faecalis</i> subjected to 0.25×, 0.5×, and 0.75× of the minimal inhibitory concentration (MIC) of ampicillin on the expression of an aggregation substance (AS) by a donor plasmid clone (1390R). A clumping assay was performed. The relative expression of <i>prgB</i> (gene that encodes AS) was determined and semiquantified in 1390R, and <i>iad1</i> expression was determined and semiquantified in C29. AS expression was analyzed in the stimulated 1390R cells by confocal microscopy, flow cytometry, and ELISA. Adherence was also measured. Maximal clumping was observed with the pheromone medium 0.25×. Only the 1390R strain stimulated with the C29 supernatant without ampicillin and with 0.25× was able to express <i>prgB</i>. No expression of <i>prgB</i> was observed at 0.5× and 0.75×. The difference in relative expression (RE) of 1390R without ampicillin and with 0.25× was 0.5-fold. AS expression in 1390R showed the greatest increase upon stimulation with 0.25×. When 1390R was stimulated with 0.5× and 0.75×, AS expression was also observed but was significantly lower. Ampicillin stimulated C29 switch-off pheromone expression in recipient cells, which in turn switched off AS expression in donor cells. We observed that although <i>prgB</i> was switched off after 0.5× stimulation in C29, the supernatants induced expression in certain 1390R strains. In conclusion, ampicillin was able to modulate pheromone expression in free plasmid clones which, in turn, modulated AS expression in plasmid donor cells. The fact that PrgB gene expression was switched off after the ampicillin stimulus at 0.5× MIC, whereas AS proteins were present on the surface of the bacteria, suggested that a mechanism of rescue associated with mechanism pheromone sensing may be involved. |
| first_indexed | 2024-03-09T18:08:25Z |
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| id | doaj.art-ee97b2161d0c4b66a54de84d94f05522 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T18:08:25Z |
| publishDate | 2022-11-01 |
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| series | Microorganisms |
| spelling | doaj.art-ee97b2161d0c4b66a54de84d94f055222023-11-24T09:17:59ZengMDPI AGMicroorganisms2076-26072022-11-011011229410.3390/microorganisms10112294Pheromone Activity after Stimulation with Ampicillin in a Plasmid-Free <i>Enterococcus faecalis</i> StrainJosé Arellano-Galindo0Sergio Zavala-Vega1Rosario Vázquez-Larios2Sara A. Ochoa3Ariadnna Cruz-Córdova4Adolfo Sierra-Santoyo5Lourdes López-González6Rigoberto Hernández-Castro7Silvia Giono-Cerezo8Juan Xicohtencatl-Cortes9Laboratorio de Bacteriología Médica, Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Ciudad de México 11340, MexicoLaboratorio de Neuropatología, Instituto Nacional de Neurología y Neurocirugía Manuel Velasco Suárez, Ciudad de México 14269, MexicoDepartamento de Microbiología, Instituto Nacional de Cardiología Ignacio Chavez, Ciudad de México 14080, MexicoLaboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Ciudad de México 06720, MexicoLaboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Ciudad de México 06720, MexicoLaboratorio de Toxicología de Plaguicidas y Disrupción Endócrina, CINVESTAV IPN, Ciudad de México 07360, MexicoLaboratorio de Toxicología de Plaguicidas y Disrupción Endócrina, CINVESTAV IPN, Ciudad de México 07360, MexicoDepartamento de Ecología de Agentes Patógenos, Hospital General Dr. Manuel Gea González, Ciudad de México 14080, MexicoLaboratorio de Bacteriología Médica, Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Ciudad de México 11340, MexicoLaboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Ciudad de México 06720, MexicoEnterococci exhibit clumping under the selective pressure of antibiotics. The aim of this study was to analyze the effect of supernatants from a plasmid-free clone (C29) of <i>Enterococcus faecalis</i> subjected to 0.25×, 0.5×, and 0.75× of the minimal inhibitory concentration (MIC) of ampicillin on the expression of an aggregation substance (AS) by a donor plasmid clone (1390R). A clumping assay was performed. The relative expression of <i>prgB</i> (gene that encodes AS) was determined and semiquantified in 1390R, and <i>iad1</i> expression was determined and semiquantified in C29. AS expression was analyzed in the stimulated 1390R cells by confocal microscopy, flow cytometry, and ELISA. Adherence was also measured. Maximal clumping was observed with the pheromone medium 0.25×. Only the 1390R strain stimulated with the C29 supernatant without ampicillin and with 0.25× was able to express <i>prgB</i>. No expression of <i>prgB</i> was observed at 0.5× and 0.75×. The difference in relative expression (RE) of 1390R without ampicillin and with 0.25× was 0.5-fold. AS expression in 1390R showed the greatest increase upon stimulation with 0.25×. When 1390R was stimulated with 0.5× and 0.75×, AS expression was also observed but was significantly lower. Ampicillin stimulated C29 switch-off pheromone expression in recipient cells, which in turn switched off AS expression in donor cells. We observed that although <i>prgB</i> was switched off after 0.5× stimulation in C29, the supernatants induced expression in certain 1390R strains. In conclusion, ampicillin was able to modulate pheromone expression in free plasmid clones which, in turn, modulated AS expression in plasmid donor cells. The fact that PrgB gene expression was switched off after the ampicillin stimulus at 0.5× MIC, whereas AS proteins were present on the surface of the bacteria, suggested that a mechanism of rescue associated with mechanism pheromone sensing may be involved.https://www.mdpi.com/2076-2607/10/11/2294pheromonesaggregation substancesampicillinantimicrobial resistanceplasmid |
| spellingShingle | José Arellano-Galindo Sergio Zavala-Vega Rosario Vázquez-Larios Sara A. Ochoa Ariadnna Cruz-Córdova Adolfo Sierra-Santoyo Lourdes López-González Rigoberto Hernández-Castro Silvia Giono-Cerezo Juan Xicohtencatl-Cortes Pheromone Activity after Stimulation with Ampicillin in a Plasmid-Free <i>Enterococcus faecalis</i> Strain Microorganisms pheromones aggregation substances ampicillin antimicrobial resistance plasmid |
| title | Pheromone Activity after Stimulation with Ampicillin in a Plasmid-Free <i>Enterococcus faecalis</i> Strain |
| title_full | Pheromone Activity after Stimulation with Ampicillin in a Plasmid-Free <i>Enterococcus faecalis</i> Strain |
| title_fullStr | Pheromone Activity after Stimulation with Ampicillin in a Plasmid-Free <i>Enterococcus faecalis</i> Strain |
| title_full_unstemmed | Pheromone Activity after Stimulation with Ampicillin in a Plasmid-Free <i>Enterococcus faecalis</i> Strain |
| title_short | Pheromone Activity after Stimulation with Ampicillin in a Plasmid-Free <i>Enterococcus faecalis</i> Strain |
| title_sort | pheromone activity after stimulation with ampicillin in a plasmid free i enterococcus faecalis i strain |
| topic | pheromones aggregation substances ampicillin antimicrobial resistance plasmid |
| url | https://www.mdpi.com/2076-2607/10/11/2294 |
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