Studi Awal: Identifikasi Keragaman Bakteri Saliva Wanita Usia Lanjut Menggunakan Teknologi 16S rRNA Sequencing Oxford Nanopore
DOI:
https://doi.org/10.15408/pbsj.v7i1.49111Keywords:
16S rRNA, Epi2Me, ONT, oral bacteria, elderly womanAbstract
The elderly are a population particularly susceptible to diseases resulting from the decline in immune system function. The mouth is one of the areas of the body that is the entry point for microorganisms into the body. It is known that the oral microbiome greatly affects oral and systemic health. The oral bacterial profile is important to understand its role in maintaining health and causing disease. Therefore, maintaining oral health is very important, especially in our study of the elderly. Utilizing advanced sequencing techniques such as Oxford Nanopore Technology (ONT) offers the opportunity to reveal bacterial diversity comprehensively. This study aims to identify the oral microbiome in a group of elderly women. Knowledge of the oral microbiota profile could help spread awareness of the benefits of maintaining good dental hygiene, especially in older adults. Methods: Seven elderly housewives had their unstimulated saliva samples taken in the morning and kept at -80°C. The phenol-chloroform technique was used to isolate the DNA, and a Qubit Fluorometer was used to measure purity and concentration. By employing ONT for 16S rRNA sequencing and Epi2Me software for analysis, taxonomic mapping of the bacterial diversity was accomplished. Results: According to the taxonomic analysis, Streptococcus and Veillonella were the most common genera in the samples. It was common to find species like Veillonella parvula, Veillonella atypica, Streptococcus mitis, and Streptococcus rubneri. These genera contributed to the dynamics of oral biofilms by exhibiting complex ecological interactions. With advanced sequencing technologies, this work highlights the diversity of oral bacteria in elderly women in Indonesia while providing a new perspective. Notwithstanding a few limitations, these findings provide insight into the ecological dynamics of the oral microbiota, potentially leading to clinical applications in the treatment and prevention of disease.
References
Abranches, J., Zeng, L et al. (2018). Biology of Oral Streptococci. Microbiology spectrum, 6(5):10.1128/microbiolspec.GPP3-0042-2018. https://doi.org/10.1128/microbiolspec.GPP3-0042-2018
Adhiyanto, C., Hendarmin, L. A., Puspitaningrum, R. Pengenalan Dasar Teknik Bio-Molekuler (Hendarto H, ed.). Sleman. Deepublish Press,2020. ISBN. 978-623-02-1505-6
Baker, J. L., Mark Welch, J. L et al. (2023). The oral microbiome: diversity, biogeography and human health. Nature reviews. Microbiology, 22: 89- 104. Advanced online publication. https://doi.org/10.1038/s41579-023-00963-6
Benn, A., Heng, N et al. (2018). Studying the human oral microbiome: Challenges and the evolution of solutions. Australian Dental Journal, 63(1): 14-24. https://doi.org/10.1111/adj.12565
Buetas, E., Jordán-López, M., López-Roldán, A., Martínez-Priego, L., Marco, G. D., Carda-Diéguez, M., Mira, A. (2024). Full-length 16S rRNA gene sequencing by PacBio improves taxonomic resolution in human microbiome samples. BMC Genomics, 25, 310. https://doi.org/10.1186/s12864-024-10213-5
Deo, P.N., Deshmukh, R. (2019). Oral microbiome: Unveiling the fundamentals. Journal of oral and maxillofacial pathology. Journal of Oral and Maxillofacial Pathology, 23(1):122–128. https://doi.org/10.4103/jomfp.JOMFP_304_18
Di Giulio, M., di Giacomo, V et al. (2013). Saliva improves Streptococcus mitis protective effect on human gingival fibroblasts in presence of 2-hydroxyethyl-methacrylate. Journal of materials science. Materials in medicine, 24(8):1977–1983. https://doi.org/10.1007/s10856-013-4949-7
Djais, A.A., Theodorea, C. F et al. (2019). Identification and phylogenetic analysis of oral Veillonella species isolated from the saliva of Japanese children. F1000Research, 8:616. https://doi.org/10.12688/f1000research.18506.5
Giacomini, J. J., Torres-Morales, J et al. (2023). Site Specialization of Human Oral Veillonella Species. ASM Journal,11(1):1-16. DOI: https://doi.org/10.1128/spectrum.04042-22
Goodwin S, McPherson JD, McCombie WR. Coming of age: ten years of next-generation sequencing technologies. Nat Rev Genet. 2016;17(6):333–351. doi:10.1038/nrg.2016.49
Hendrawan, R.B. (2024). Efek Negatif Merokok terhadap Kesehatan Gigi dan Gusi. https://fkg.ugm.ac.id/id/efek-negatif-merokok-terhadap-kesehatan-gigi-dan-gusi/ diakses: 24 Juni 2025
Heng, N. C. K., Stanton, J. A. L. (2019). Next-generation DNA sequencing of oral microbes at the Sir John Walsh Research Institute: technologies, tools, and achievements. Journal of the Royal Society of New Zealand, 50(1), 91-107. https://doi.org/10.1080/03036758.2019.1687530
Huch, M., De Bruyne, K et al. (2013). Streptococcus rubneri sp. nov., isolated from the human throat. International journal of systematic and evolutionary microbiology, 63(11):4026–4032. https://doi.org/10.1099/ijs.0.048538-0
Kilian, M., Chapple, I. L et al. (2016). The oral microbiome – an update for oral healthcare professionals. British Dental Journal, 221(10): 657-666. https://doi.org/10.1038/sj.bdj.2016.865
Leake, S. L., Pagni, M et al. (2016). The salivary microbiome for differentiating individuals: Proof of principle. Microbes and Infection, 18(6): 399-405. https://doi.org/10.1016/j.micinf.2016.03.011
Lu, M., Xuan, S., Wang, Z. (2019). Oral microbiota: A new view of body health. Food Science and Human Wellness, 8(1):8-15. https://doi.org/10.1016/j.fshw.2018.12.001
Lu, Hongye, Peihui Zou, Yifei Zhang, Qian Zhang, Zhibin Chen, and Feng Chen. 2022. “ The Sampling Strategy of Oral Microbiome.” iMeta 1, e23. https://doi.org/10.1002/imt2.23
Maheshwari, K., Musyuni, P., Moulick, A., Mishra, H., Ekielski, A., Mishra, P.K., Aggarwal, G. (2024). Unveiling the microbial symphony: Next-Gen sequencing and bioinformatics insights into the human gut microbiome. Health Sciences Review, 11, 100173. https://doi.org/10.1016/j.hsr.2024.100173
Morrison, A.G., Sarkar, S., Umar, S., Lee, S.T., Thomas, S. M. (2023). The Contribution of the Human Oral Microbiome to Oral Disease: A Review. Microorganisms,11(2), 318. https://doi.org/10.3390/microorganisms11020318
Miller MB, Tang YW. Basic concepts of microarrays and potential applications in clinical microbiology. Clin Microbiol Rev. 2009;22(4):611–633. doi:10.1128/CMR.00019-09
N/A- (2023). What does smoking do to oral bacteria? https://www.dental-nursing.co.uk/news/what-does-smoking-do-to-oral-bacteria diakses: 24 Juni 2025
Oxford Nanopore Technology. https://community.nanoporetech.com/docs/prepare/library_prep_protocols/rapid-sequencing-DNA-16s-barcoding-kit-v14-sqk-16114-24/v/16s_9199_v114_reva_06dec2023
Pathak JL, Yan Y, Zhang Q, Wang L, Ge L. (2021). The role of oral microbiome in respiratory health and diseases. Respiratory Medicine, 185. 106475. https://doi.org/10.1016/j.rmed.2021.106475
Peng, X., Cheng, L et al. (2022). Oral microbiota in human systematic diseases. International Journal of Oral Science, 14(1): 1-11. https://doi.org/10.1038/s41368-022-00163-7
Pisano, Massimo et al. (2023). The Interaction between the Oral Microbiome and Systemic Diseases: A Narrative Review. Microbiology Research, 14(4): 1862-1878. https://doi.org/10.3390/microbiolres14040127
Pozhitkov, A. E., Beikler, T., Flemmig, T., Noble, P.A. (2011). High-throughput methods for analysis of the human oral microbiome. Periodontology 2000, 55(1): 70–86. https://doi.org/10.1111/j.1600-0757.2010.00380.x
Rajasekaran, J. J., Krishnamurthy, H. K., Bosco, J., Jayaraman, V., Krishna, K., Wang, T., Bei, K. (2024). Oral Microbiome: A Review of Its Impact on Oral and Systemic Health. Microorganisms, 12(9): 1797. https://doi.org/10.3390/microorganisms12091797
Santacroce, L, Passarelli et al. (2023). Oral microbiota in human health and disease: A perspective. Experimental Biology and Medicine, 248(15):1288-1301. https://doi:10.1177/15353702231187645
Satam, H., Joshi, K., Mangrolia, U., Waghoo, S., Zaidi, G., Rawool, S., Thakare, R. P., Banday, S., Mishra, A.K., Das, G., Malonia, S.K. (2023). Next-Generation Sequencing Technology: Current Trends and Advancements. Biology, 12(7), 997. https://doi.org/10.3390/biology12070997
Schwarz, C., Hajdu, A.I et al. (2023). Link between Oral Health, Periodontal Disease, Smoking, and Systemic Diseases in Romanian Patients. Healthcare, 11(16):2354 https://doi.org/10.3390/healthcare11162354
Sedghi, L., DiMassa, V., Harrington, A., Lynch, S.V., Kapila, Y.L. (2021).The oral microbiome: Role of key organisms and complex networks in oral health and disease. Periodontology 2000, 87(1):107-131. https://doi.org/10.1111/prd.12393
Wang, J., Feng, J et al. (2022). Diversity and Biogeography of Human Oral Saliva Microbial Communities Revealed by the Earth Microbiome Project. Frontiers in Microbiology, 13:931065. https://doi.org/10.3389/fmicb.2022.931065
Widyarman, A. S., Theodorea, C.F., Udawatte, N.S., Drestia, A.M., Bachtiar, E.W., Astoeti, T. E., Bachtiar, B.M. (2021). Diversity of Oral Microbiome of Women From Urban and Rural Areas of Indonesia: A Pilot Study. Frontiers in Oral Health, 2, 738306. https://doi.org/10.3389/froh.2021.738306
Willis, J. R., Gabaldón, T. (2020). The Human Oral Microbiome in Health and Disease: From Sequences to Ecosystems. Microorganisms, 8(2):308 https://doi.org/10.3390/microorganisms8020308
Willenborg, J., Goethe, R. (2016). Metabolic traits of pathogenic streptococci. FEBS Letters, 590(21): 3905-3919. https://doi.org/10.1002/1873-3468.12317
Zhou, P., Liu, J et al. (2015). A YadA-like autotransporter, Hag1 in Veillonella atypica is a multivalent hemagglutinin involved in adherence to oral streptococci, Porphyromonas gingivalis, and human oral buccal cells. Molecular oral microbiology, 30(4): 269–279. https://doi.org/10.1111/omi.12091
Zhou, P., Manoil, D et al. (2021). Veillonellae: Beyond Bridging Species in Oral Biofilm Ecology. Frontiers in oral health, 2:774115. https://doi.org/10.3389/froh.2021.774115
Downloads
Published
Issue
Section
License
Copyright (c) 2025 Chris Adhiyanto., Ph.D, Laifa A. Hendarmin, Zeti Harriyati, Erike A. Suwarsono, Auliyani Andam Suri , Yudhi Nugraha, Dwirini Retno Gunarti

This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.