Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase Activity
Streptococcus pneumoniae colonizes the upper airways of children and the elderly. Colonization progresses to persistent carriage when S. pneumoniae forms biofilms, a feature required for the development of pneumococcal disease. Nasopharyngeal biofilms are structured with a matrix that includes extra...
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| Format: | Article |
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Frontiers Media S.A.
2019-10-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.02386/full |
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| author | Uriel A. Angulo-Zamudio Uriel A. Angulo-Zamudio Jorge E. Vidal Jorge E. Vidal Kamran Nazmi Jan G. M. Bolscher Claudia Leon-Sicairos Brenda S. Antezana Adrián Canizalez-Roman Adrián Canizalez-Roman Nidia León-Sicairos Nidia León-Sicairos |
| author_facet | Uriel A. Angulo-Zamudio Uriel A. Angulo-Zamudio Jorge E. Vidal Jorge E. Vidal Kamran Nazmi Jan G. M. Bolscher Claudia Leon-Sicairos Brenda S. Antezana Adrián Canizalez-Roman Adrián Canizalez-Roman Nidia León-Sicairos Nidia León-Sicairos |
| author_sort | Uriel A. Angulo-Zamudio |
| collection | DOAJ |
| description | Streptococcus pneumoniae colonizes the upper airways of children and the elderly. Colonization progresses to persistent carriage when S. pneumoniae forms biofilms, a feature required for the development of pneumococcal disease. Nasopharyngeal biofilms are structured with a matrix that includes extracellular DNA (eDNA), which is sourced from the same pneumococci and other bacteria. This eDNA also allows pneumococci to acquire new traits, including antibiotic resistance genes. In this study, we investigated the efficacy of lactoferrin (LF), at physiological concentrations found in secretions with bactericidal activity [i.e., colostrum (100 μM), tears (25 μM)], in eradicating pneumococcal biofilms from human respiratory cells. The efficacy of synthetic LF-derived peptides was also assessed. We first demonstrated that LF inhibited colonization of S. pneumoniae on human respiratory cells without affecting the viability of planktonic bacteria. LF-derived peptides were, however, bactericidal for planktonic pneumococci but they did not affect viability of pre-formed biofilms. In contrast, LF (40 and 80 μM) eradicated pneumococcal biofilms that had been pre-formed on abiotic surfaces (i.e., polystyrene) and on human pharyngeal cells, as investigated by viable counts and confocal microscopy. LF also eradicated biofilms formed by S. pneumoniae strains with resistance to multiple antibiotics. We investigated whether treatment with LF would affect the biofilm structure by analyzing eDNA. Surprisingly, in pneumococcal biofilms treated with LF, the eDNA was absent in comparison to the untreated control (∼10 μg/ml) or those treated with LF-derived peptides. EMSA assays showed that LF binds S. pneumoniae DNA and a time-course study of DNA decay demonstrated that the DNA is degraded when bound by LF. This LF-associated DNase activity inhibited acquisition of antibiotic resistance genes in both in vitro transformation assays and in a life-like bioreactor system. In conclusion, we demonstrated that LF eradicates pneumococcal-colonizing biofilms at a concentration safe for humans and identified a LF-associated DNAse activity that inhibited the acquisition of resistance. |
| first_indexed | 2024-12-17T12:50:52Z |
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| language | English |
| last_indexed | 2024-12-17T12:50:52Z |
| publishDate | 2019-10-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-019236c317f24226939404b15ecaeb8e2022-12-21T21:47:36ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-10-011010.3389/fmicb.2019.02386492686Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase ActivityUriel A. Angulo-Zamudio0Uriel A. Angulo-Zamudio1Jorge E. Vidal2Jorge E. Vidal3Kamran Nazmi4Jan G. M. Bolscher5Claudia Leon-Sicairos6Brenda S. Antezana7Adrián Canizalez-Roman8Adrián Canizalez-Roman9Nidia León-Sicairos10Nidia León-Sicairos11CIASaP, Facultad de Medicina, Universidad Autónoma de Sinaloa, Culiacán, MexicoPrograma Regional del Noroeste para el Doctorado en Biotecnología, Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Sinaloa, Culiacán, MexicoHubert Department of Global Health, Rollins School of Public Health, Emory University, Atlanta, GA, United StatesDepartment of Microbiology and Immunology, The University of Mississippi Medical Center, Jackson, MS, United StatesDepartment of Oral Biochemistry, Academic Centre for Dentistry Amsterdam, University of Amsterdam, Amsterdam, NetherlandsDepartment of Oral Biochemistry, Academic Centre for Dentistry Amsterdam, University of Amsterdam, Amsterdam, NetherlandsPrograma Regional del Noroeste para el Doctorado en Biotecnología, Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Sinaloa, Culiacán, MexicoMicrobiology and Molecular Genetics Program, Graduate Division of Biological and Biomedical Sciences, Emory University, Atlanta, GA, United StatesCIASaP, Facultad de Medicina, Universidad Autónoma de Sinaloa, Culiacán, MexicoUnidad de Investigación, Hospital de la Mujer, Servicios de Salud de Sinaloa, Culiacán, MexicoCIASaP, Facultad de Medicina, Universidad Autónoma de Sinaloa, Culiacán, MexicoDepartamento de Investigación del Hospital Pediátrico de Sinaloa, Servicios de Salud de Sinaloa, Culiacán, MexicoStreptococcus pneumoniae colonizes the upper airways of children and the elderly. Colonization progresses to persistent carriage when S. pneumoniae forms biofilms, a feature required for the development of pneumococcal disease. Nasopharyngeal biofilms are structured with a matrix that includes extracellular DNA (eDNA), which is sourced from the same pneumococci and other bacteria. This eDNA also allows pneumococci to acquire new traits, including antibiotic resistance genes. In this study, we investigated the efficacy of lactoferrin (LF), at physiological concentrations found in secretions with bactericidal activity [i.e., colostrum (100 μM), tears (25 μM)], in eradicating pneumococcal biofilms from human respiratory cells. The efficacy of synthetic LF-derived peptides was also assessed. We first demonstrated that LF inhibited colonization of S. pneumoniae on human respiratory cells without affecting the viability of planktonic bacteria. LF-derived peptides were, however, bactericidal for planktonic pneumococci but they did not affect viability of pre-formed biofilms. In contrast, LF (40 and 80 μM) eradicated pneumococcal biofilms that had been pre-formed on abiotic surfaces (i.e., polystyrene) and on human pharyngeal cells, as investigated by viable counts and confocal microscopy. LF also eradicated biofilms formed by S. pneumoniae strains with resistance to multiple antibiotics. We investigated whether treatment with LF would affect the biofilm structure by analyzing eDNA. Surprisingly, in pneumococcal biofilms treated with LF, the eDNA was absent in comparison to the untreated control (∼10 μg/ml) or those treated with LF-derived peptides. EMSA assays showed that LF binds S. pneumoniae DNA and a time-course study of DNA decay demonstrated that the DNA is degraded when bound by LF. This LF-associated DNase activity inhibited acquisition of antibiotic resistance genes in both in vitro transformation assays and in a life-like bioreactor system. In conclusion, we demonstrated that LF eradicates pneumococcal-colonizing biofilms at a concentration safe for humans and identified a LF-associated DNAse activity that inhibited the acquisition of resistance.https://www.frontiersin.org/article/10.3389/fmicb.2019.02386/fulllactoferrindisaggregatespneumococcal biofilmsDNAse activityresistance acquisition |
| spellingShingle | Uriel A. Angulo-Zamudio Uriel A. Angulo-Zamudio Jorge E. Vidal Jorge E. Vidal Kamran Nazmi Jan G. M. Bolscher Claudia Leon-Sicairos Brenda S. Antezana Adrián Canizalez-Roman Adrián Canizalez-Roman Nidia León-Sicairos Nidia León-Sicairos Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase Activity Frontiers in Microbiology lactoferrin disaggregates pneumococcal biofilms DNAse activity resistance acquisition |
| title | Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase Activity |
| title_full | Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase Activity |
| title_fullStr | Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase Activity |
| title_full_unstemmed | Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase Activity |
| title_short | Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase Activity |
| title_sort | lactoferrin disaggregates pneumococcal biofilms and inhibits acquisition of resistance through its dnase activity |
| topic | lactoferrin disaggregates pneumococcal biofilms DNAse activity resistance acquisition |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2019.02386/full |
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