The effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiome
Abstract Background Oral microbiome played an important role in maintaining healthy state and might exhibit certain changes under circumstances of diseases. However, current microbiological research using sequencing techniques did not regard dead bacteria as a separate part, causing findings based o...
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| Format: | Article |
| Language: | English |
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BMC
2021-09-01
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| Series: | BMC Oral Health |
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| Online Access: | https://doi.org/10.1186/s12903-021-01832-5 |
| _version_ | 1818589410428452864 |
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| author | Qidi Ren Fangqiao Wei Chao Yuan Ce Zhu Qian Zhang Junkang Quan Xiangyu Sun Shuguo Zheng |
| author_facet | Qidi Ren Fangqiao Wei Chao Yuan Ce Zhu Qian Zhang Junkang Quan Xiangyu Sun Shuguo Zheng |
| author_sort | Qidi Ren |
| collection | DOAJ |
| description | Abstract Background Oral microbiome played an important role in maintaining healthy state and might exhibit certain changes under circumstances of diseases. However, current microbiological research using sequencing techniques did not regard dead bacteria as a separate part, causing findings based on subsequent analyses on dynamic equilibrium and functional pathways of microbes somewhat questionable. Since treatment by propidium monoazide (PMA) was able to remove dead bacteria effectively, it would be worth studying how the sequencing results after PMA treatment differed from those focusing on the whole microbiota. Methods Unstimulated whole saliva samples were obtained from 18 healthy people from 3 age groups (children, adults, and the elderly). After removal of dead bacteria by propidium monoazide (PMA), changes in the profile of salivary microbiome were detected using 16S rRNA sequencing technology, and differences among age groups were compared subsequently. Results Dead bacteria accounted for nearly a half of the whole bacteria flora in saliva, while freezing had little effect on the proportion of deaths. After treatment with PMA, the numbers of OTUs reduced by 4.4–14.2%, while the Shannon diversity indices decreased significantly (P < 0.01). Only 35.2% of positive and 6.1% of negative correlations were found to be shared by the whole microbiota and that with dead bacteria removed. Differences in significantly changed OTUs and functional pathways among different age groups were also observed between the group of PMA and the control. Conclusions It was necessary to take the influence of living state of bacteria into account in analytic studies of salivary microbiome. |
| first_indexed | 2024-12-16T09:40:12Z |
| format | Article |
| id | doaj.art-1a35fb2067674b39ac7c270882dfb115 |
| institution | Directory Open Access Journal |
| issn | 1472-6831 |
| language | English |
| last_indexed | 2024-12-16T09:40:12Z |
| publishDate | 2021-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Oral Health |
| spelling | doaj.art-1a35fb2067674b39ac7c270882dfb1152022-12-21T22:36:18ZengBMCBMC Oral Health1472-68312021-09-0121111210.1186/s12903-021-01832-5The effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiomeQidi Ren0Fangqiao Wei1Chao Yuan2Ce Zhu3Qian Zhang4Junkang Quan5Xiangyu Sun6Shuguo Zheng7Department of Preventive Dentistry, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of StomatologyDepartment of Preventive Dentistry, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of StomatologyDepartment of Preventive Dentistry, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of StomatologyDepartment of Preventive Dentistry, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of StomatologyCentral Laboratory, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of StomatologyDepartment of Preventive Dentistry, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of StomatologyDepartment of Preventive Dentistry, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of StomatologyDepartment of Preventive Dentistry, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of StomatologyAbstract Background Oral microbiome played an important role in maintaining healthy state and might exhibit certain changes under circumstances of diseases. However, current microbiological research using sequencing techniques did not regard dead bacteria as a separate part, causing findings based on subsequent analyses on dynamic equilibrium and functional pathways of microbes somewhat questionable. Since treatment by propidium monoazide (PMA) was able to remove dead bacteria effectively, it would be worth studying how the sequencing results after PMA treatment differed from those focusing on the whole microbiota. Methods Unstimulated whole saliva samples were obtained from 18 healthy people from 3 age groups (children, adults, and the elderly). After removal of dead bacteria by propidium monoazide (PMA), changes in the profile of salivary microbiome were detected using 16S rRNA sequencing technology, and differences among age groups were compared subsequently. Results Dead bacteria accounted for nearly a half of the whole bacteria flora in saliva, while freezing had little effect on the proportion of deaths. After treatment with PMA, the numbers of OTUs reduced by 4.4–14.2%, while the Shannon diversity indices decreased significantly (P < 0.01). Only 35.2% of positive and 6.1% of negative correlations were found to be shared by the whole microbiota and that with dead bacteria removed. Differences in significantly changed OTUs and functional pathways among different age groups were also observed between the group of PMA and the control. Conclusions It was necessary to take the influence of living state of bacteria into account in analytic studies of salivary microbiome.https://doi.org/10.1186/s12903-021-01832-5Salivary microbiomePropidium monoazideRemoval of dead bacteria16S rRNA sequencing |
| spellingShingle | Qidi Ren Fangqiao Wei Chao Yuan Ce Zhu Qian Zhang Junkang Quan Xiangyu Sun Shuguo Zheng The effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiome BMC Oral Health Salivary microbiome Propidium monoazide Removal of dead bacteria 16S rRNA sequencing |
| title | The effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiome |
| title_full | The effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiome |
| title_fullStr | The effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiome |
| title_full_unstemmed | The effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiome |
| title_short | The effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiome |
| title_sort | effects of removing dead bacteria by propidium monoazide on the profile of salivary microbiome |
| topic | Salivary microbiome Propidium monoazide Removal of dead bacteria 16S rRNA sequencing |
| url | https://doi.org/10.1186/s12903-021-01832-5 |
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