Carbonyl Cyanide 3-Chloro Phenyl Hydrazone (CCCP) Restores the Colistin Sensitivity in <i>Brucella intermedia</i>
<i>Brucella intermedia</i> (formerly <i>Ochrobactrum intermedium</i>), a non-fermentative bacterium, has been isolated from animals and human clinical specimens. It is naturally resistant to polymyxins, including colistin (CO), and may cause opportunistic infections in humans...
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MDPI AG
2023-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/3/2106 |
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| author | Malak Zoaiter Zaher Zeaiter Oleg Mediannikov Cheikh Sokhna Pierre-Edouard Fournier |
| author_facet | Malak Zoaiter Zaher Zeaiter Oleg Mediannikov Cheikh Sokhna Pierre-Edouard Fournier |
| author_sort | Malak Zoaiter |
| collection | DOAJ |
| description | <i>Brucella intermedia</i> (formerly <i>Ochrobactrum intermedium</i>), a non-fermentative bacterium, has been isolated from animals and human clinical specimens. It is naturally resistant to polymyxins, including colistin (CO), and may cause opportunistic infections in humans. We isolated six <i>Brucella intermedia</i> strains from Senegalese monkey stool. In order to determine whether an efflux pump mechanism was involved in CO resistance in <i>B. intermedia,</i> we evaluated the effects of verapamil (VRP), reserpine (RSP), phe-arg β-naphthylamide dihydrochloride (PAβN) and carbonyl cyanide 3-chloro phenyl hydrazone (CCCP), four efflux pump inhibitors, on these colistin-resistant strains. Using the broth microdilution method, a CO and CCCP combination of 2 µg/mL and 10 µg/mL, respectively, significantly reduced the CO minimal inhibitory concentration (MIC) of <i>B. intermedia</i>, supporting an efflux pump mechanism. In contrast, VRP, PAβN and RSP did not restore CO susceptibility. A time kill assay showed a bactericidal effect of the CO–CCCP combination. Genomic analysis revealed a potential implication in the CO resistance mechanism of some conserved efflux pumps, such as YejABEF, NorM and EmrAB, as previously reported in other bacteria. An inhibitory effect of the CO–CCCP combination was observed on biofilm formation using the crystal violet method. These results suggest that the intrinsic CO resistance in <i>Brucella intermedia</i> is linked to an efflux pump mechanism and that the synergistic effect of CO–CCCP may open a new field to identify new treatments to restore antibiotic efficacy in humans. |
| first_indexed | 2024-03-11T09:42:30Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T09:42:30Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-13f82c03c15746b3ae1015a10062f5912023-11-16T16:52:26ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-01-01243210610.3390/ijms24032106Carbonyl Cyanide 3-Chloro Phenyl Hydrazone (CCCP) Restores the Colistin Sensitivity in <i>Brucella intermedia</i>Malak Zoaiter0Zaher Zeaiter1Oleg Mediannikov2Cheikh Sokhna3Pierre-Edouard Fournier4Institut Hospitalo-Universitaire Méditerranée-Infection, 13005 Marseille, FranceDepartment of Biology, Faculty of Sciences, Lebanese University LU, Beirut 146404, LebanonInstitut Hospitalo-Universitaire Méditerranée-Infection, 13005 Marseille, FranceInstitut Hospitalo-Universitaire Méditerranée-Infection, 13005 Marseille, FranceInstitut Hospitalo-Universitaire Méditerranée-Infection, 13005 Marseille, France<i>Brucella intermedia</i> (formerly <i>Ochrobactrum intermedium</i>), a non-fermentative bacterium, has been isolated from animals and human clinical specimens. It is naturally resistant to polymyxins, including colistin (CO), and may cause opportunistic infections in humans. We isolated six <i>Brucella intermedia</i> strains from Senegalese monkey stool. In order to determine whether an efflux pump mechanism was involved in CO resistance in <i>B. intermedia,</i> we evaluated the effects of verapamil (VRP), reserpine (RSP), phe-arg β-naphthylamide dihydrochloride (PAβN) and carbonyl cyanide 3-chloro phenyl hydrazone (CCCP), four efflux pump inhibitors, on these colistin-resistant strains. Using the broth microdilution method, a CO and CCCP combination of 2 µg/mL and 10 µg/mL, respectively, significantly reduced the CO minimal inhibitory concentration (MIC) of <i>B. intermedia</i>, supporting an efflux pump mechanism. In contrast, VRP, PAβN and RSP did not restore CO susceptibility. A time kill assay showed a bactericidal effect of the CO–CCCP combination. Genomic analysis revealed a potential implication in the CO resistance mechanism of some conserved efflux pumps, such as YejABEF, NorM and EmrAB, as previously reported in other bacteria. An inhibitory effect of the CO–CCCP combination was observed on biofilm formation using the crystal violet method. These results suggest that the intrinsic CO resistance in <i>Brucella intermedia</i> is linked to an efflux pump mechanism and that the synergistic effect of CO–CCCP may open a new field to identify new treatments to restore antibiotic efficacy in humans.https://www.mdpi.com/1422-0067/24/3/2106intrinsic colistin resistanceefflux pump inhibitorscarbonyl cyanide 3-chloro phenyl hydrazoneCO-MICsynergyefflux pump |
| spellingShingle | Malak Zoaiter Zaher Zeaiter Oleg Mediannikov Cheikh Sokhna Pierre-Edouard Fournier Carbonyl Cyanide 3-Chloro Phenyl Hydrazone (CCCP) Restores the Colistin Sensitivity in <i>Brucella intermedia</i> International Journal of Molecular Sciences intrinsic colistin resistance efflux pump inhibitors carbonyl cyanide 3-chloro phenyl hydrazone CO-MIC synergy efflux pump |
| title | Carbonyl Cyanide 3-Chloro Phenyl Hydrazone (CCCP) Restores the Colistin Sensitivity in <i>Brucella intermedia</i> |
| title_full | Carbonyl Cyanide 3-Chloro Phenyl Hydrazone (CCCP) Restores the Colistin Sensitivity in <i>Brucella intermedia</i> |
| title_fullStr | Carbonyl Cyanide 3-Chloro Phenyl Hydrazone (CCCP) Restores the Colistin Sensitivity in <i>Brucella intermedia</i> |
| title_full_unstemmed | Carbonyl Cyanide 3-Chloro Phenyl Hydrazone (CCCP) Restores the Colistin Sensitivity in <i>Brucella intermedia</i> |
| title_short | Carbonyl Cyanide 3-Chloro Phenyl Hydrazone (CCCP) Restores the Colistin Sensitivity in <i>Brucella intermedia</i> |
| title_sort | carbonyl cyanide 3 chloro phenyl hydrazone cccp restores the colistin sensitivity in i brucella intermedia i |
| topic | intrinsic colistin resistance efflux pump inhibitors carbonyl cyanide 3-chloro phenyl hydrazone CO-MIC synergy efflux pump |
| url | https://www.mdpi.com/1422-0067/24/3/2106 |
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