Abstract
The current study aimed at the determination of the impact of obesity on the salivary microbiome in adolescents. Sixty subjects ranging 14–17 years old were enrolled (obese: n = 30–50% females, and normal weight: n = 30–50% females). Stimulated saliva was collected for denaturing gradient gel electrophoresis (DGGE) band patterns and massive 16S rRNA gene sequencing using the Ion Torrent platform. Overall, data analysis revealed that male subjects harbored a higher diverse salivary microbiome, defined by a significant higher richness (32.48 versus 26.74) and diversity (3.36 versus 3.20), higher Simpson values (0.96 versus 0.95) and distinct bacterial community structure considering either sex or condition (p < 0.05). Bacterial community fingerprinting analysis in human saliva showed a positive correlation with increased body mass index (BMI) in adolescents. Veillonella, Haemophilus and Prevotella occurrence was found to be affected by BMI, whereas Neisseria and Rothia occurrence was significantly impacted by sex in obese subjects. Our findings suggest that male and female adolescents may harbor a naturally distinct salivary microbiota and that obesity may specifically have an impact on their oral bacterial community. The potential dysbiotic oral microbiome in obese adolescents raises new insights on the etiology and prevention of future conditions in these populations.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Sequencing data are available for download at Mg-Rast (https://metagenomics.anl.gov/) under the job id—395949.
References
Błachnio-Zabielska AU, Baranowski M, Hirnle T, Zabielski P, Lewczuk A, Dmitruk I et al (2012) Increased bioactive lipids content in human subcutaneous and epicardial fat tissue correlates with insulin resistance. Lipids 47(12):1131–1141. https://doi.org/10.1007/s11745-012-3722-x
Borgo F, Verduci E, Riva A, Lassandro C, Riva E, Morace G et al (2017) Relative abundance in bacterial and fungal gut microbes in obese children: a case control study. Child Obes 13(1):78–84. https://doi.org/10.1089/chi.2015.0194
Bray JR, Curtis JT (1957) An Ordination of the upland forest communities of Southern Wisconsin. Ecol Mon 27:325–349
Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK et al (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7(5):335–336
Casadevall A (2017) The pathogenic potential of a microbe. mSphere 2:e00015–e00017. https://doi.org/10.1128/mSphere.00015-17
Cocoros NM, Lash TL, Nørgaard M, Farkas DK, DeMaria A, Sørensen HT (2013) Hospitalized prenatal and childhood infections and obesity in Danish male conscripts. Ann Epidemiol 23(6):307–313
Coretti L, Cuomo M, Florio E, Palumbo D, Keller S, Pero R et al (2017) Subgingival dysbiosis in smoker and non-smoker patients with chronic periodontitis. Mol Med Rep 15(4):2007–2014
Dalla Vecchia CF, Susin C, Rösing CK, Oppermann RV, Albandar JM (2005) Overweight and obesity as risk indicators for periodontitis in adults. J Periodontol 76(10):1721–1728. https://doi.org/10.1902/jop.2005.76.10.1721
de Onis M, Lobstein T (2010) Defining obesity risk status in the general childhood samples: which cut-offs should we use? Int J Pediatr Obes 5(6):458–460
DeGruttola AK, Low D, Mizoguchi A, Mizoguchi E (2016) Current understanding of dysbiosis in disease in human and animal models. Inflamm Bowel Dis 22(5):1137–1150
DeSantis TZ, Hugenholtz P, Larsen N, Rojas M, Brodie EL, Keller K et al (2006) Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Appl Environ Microbiol 72:5069–5072. https://doi.org/10.1128/AEM.03006-05
Dewhirst FE, Chen T, Izard J, Paster BJ, Tanner AC, Yu WH et al (2010) The human oral microbiome. J Bacteriol 192(19):5002–5017
Dhurandhar NV, Bailey D, Thomas D (2015) Interaction of obesity and infections. Obes Rev 16(12):1017–1029
DiBaise JK, Zhang H, Crowell MD, Krajmalnik-Brown R, Decker GA, Rittmann BE (2008) Gut microbiota and its possible relationship with obesity. Mayo Clin Proc 83(4):460–469. https://doi.org/10.4065/83.4.460
Ding T, Schloss PD (2014) Dynamics and associations of microbial community types across the human body. Nature 509(7500):357–360
Duran-Pinedo AE, Frias-Lopez J (2015) Beyond microbial community composition: functional activities of the oral microbiome in health and disease. Microbes Infect 17(7):505–516. https://doi.org/10.1016/j.micinf.2015.03.014
Dursun E, Akalin FA, Genc T, Cinar N, Erel O, Yildiz BO (2016) Oxidative stress and periodontal disease in obesity. Medicine (Baltimore) 95(12):e3136. https://doi.org/10.1097/MD.0000000000003136
Falagas ME, Kompoti M (2006) Obesity and infection. Lancet Infect Dis 6(7):438–446. https://doi.org/10.1016/s1473-3099(06)70523-0
Farrell JJ, Zhang L, Zhou H, Chia D, Elashoff D, Akin D et al (2012) Variations of oral microbiota are associated with pancreatic diseases including pancreatic cancer. Gut 61(4):582–588
Fierer N, Hamady M, Lauber CL, Knight R (2008) The influence of sex, handedness, and washing on the diversity of hand surface bacteria. Proc Natl Acad Sci USA 105:17994–17999. https://doi.org/10.1073/pnas.0807920105
Galvão-Moreira LV, Andrade CM, Oliveira JFF, Bomfim MRQ, Figueredo PMS, Branco-de-Almeida LS (2018) Sex differences in salivary parameters of caries susceptibility in healthy individuals. Oral Health Prev Dent. https://doi.org/10.3290/j.ohpd.a39684
Gershoni M, Pietrokovski S (2017) The landscape of sex-differential transcriptome and its consequent selection in human adults. BMC Biol 15:1–15. https://doi.org/10.1186/s12915-017-0352-z
Gevers D, Knight R, Petrosino JF, Huang K, McGuire AL, Birren BW et al (2012) The human microbiome project: a community resource for the healthy human microbiome. PLoS Biol 10:6–10. https://doi.org/10.1371/journal.pbio.1001377
Goodson JM, Groppo D, Halem S, Carpino E (2009) Is obesity an oral bacterial disease? J Dent Res 88(6):519–523. https://doi.org/10.1177/0022034509338353
Goodson JM, Hartman ML, Shi P, Hasturk H, Yaskell T, Vargas J et al (2017) The salivary microbiome is altered in the presence of a high salivary glucose concentration. PLoS One 12(3):e0170437. https://doi.org/10.1371/journal.pone.0170437
Haffajee AD, Socransky SS (2009) Relation of body mass index, periodontitis and Tannerella forsythia. J Clin Periodontol 36(2):89–99. https://doi.org/10.1111/j.1600-051X.2008.01356.x
Hainer V, Hainerová IA, Zamrazilová H (2012) Role of infection in the pathogenesis of obesity. Cas Lek Cesk 151(12):563–567
Haro C, Rangel-Zúñiga OA, Alcalá-Díaz JF, Gómez-Delgado F, Pérez-Martínez P, Delgado-Lista J et al (2016) Intestinal microbiota is influenced by gender and body mass index. PLoS One 11:1–16. https://doi.org/10.1371/journal.pone.0154090
Heuer H, Krsek M, Baker P, Smalla K, Wellington EM (1997) Analysis of actinomycete communities by specific amplification of genes encoding 16S rRNA and gel-electrophoretic separation in denaturing gradients. Appl Environ Microbiol 63(8):3233–3241
Hu HJ, Park SG, Jang HB, Min-Gyu Choi, Park KH, Kang JH et al (2015) Obesity alters the microbial community profile in korean adolescents. PLoS One 10(9):e0138015. https://doi.org/10.1371/journal.pone.0134333
Kamilar JM, Cooper N (2013) Phylogenetic signal in primate behaviour, ecology and life history. Philos Trans R Soc Lond B Biol Sci 368:20120341. https://doi.org/10.1098/rstb.2012.0341
Kopylova E, Noé L, Touzet H (2012) SortMeRNA: fast and accurate filtering of ribosomal RNAs in metatranscriptomic data. Bioinformatics 28(24):3211–3217
Kuczynski J, Stombaugh J, Walters WA, González A, Caporaso JG, Knight R (2012) Using QIIME to analyze 16S rRNA gene sequences from microbial communities. Curr Protoc Bioinform Chapter 10:Unit 10.7
Kulecka M, Paziewska A, Zeber-Lubecka N, Ambrozkiewicz F, Kopczynski M, Kuklinska U et al (2016) Prolonged transfer of feces from the lean mice modulates gut microbiota in obese mice. Nutr Metab 13(1):57. https://doi.org/10.1186/s12986-016-0116-8
Lammel DR, Barth G, Ovaskainen O, Cruz LM, Zanatta JA, Ryo M et al (2008) Direct and indirect effects of a pH gradient bring insights into the mechanisms driving prokaryotic community structures. Microbiome 6(106):7–9. https://doi.org/10.1186/s40168-018-0482-8
Li J, Quinque D, Horz H-P, Li M, Rzhetskaya M, Raff JA et al (2014) Comparative analysis of the human saliva microbiome from different climate zones: Alaska, Germany, and Africa. BMC Microbiol 14:316. https://doi.org/10.1186/s12866-014-0316-1
Li-Hui W, Chuan-Quan L, Long Y, Ru-Liu L, Long-Hui C, Wei-Wen C (2016) Gender differences in the saliva of young healthy subjects before and after citric acid stimulation. Clin Chim Acta. https://doi.org/10.1016/j.cca.2016.06.040
Liu B, Faller LL, Klitgord N, Mazumdar V, Ghodsi M, Sommer DD et al (2012) Deep sequencing of the oral microbiome reveals signatures of periodontal disease. PLoS One 7(6):e37919
Lozupone CA, Hamady M, Kelley ST, Knight R (2007) Quantitative and qualitative beta diversity measures lead to different insights into factors that structure microbial communities. Appl Environ Microbiol 73(5):1576–1585
Markle JGM, Frank DN, Mortin-toth S, Robertson CE, Feazel LM et al (2013) Sex differences in the gut. Science 80(339):1084–1088. https://doi.org/10.1126/science.1233521
McCaig AE, Glover LA, Prosser JI (2001) Numerical analysis of grassland bacterial community structure under different land management regimens by using 16S ribosomal DNA sequence data and denaturing gradient gel electrophoresis banding patterns. Appl Environ Microbiol 67(10):4554–4559
Mendes R, Raaijmakers JM (2015) Cross-kingdom similarities in microbiome functions. ISME J. https://doi.org/10.1038/ismej.2015.7
Mervish NA, Hu J, Hagan LA, Arora M, Frau C, Choi J et al (2019) Associations of the oral microbiota with obesity and menarche in inner city girls. J Child Obes 4(1):2. https://doi.org/10.21767/2572-5394.100068
Mueller S, Saunier K, Hanisch C, Norin E, Alm L, Midtvedt T et al (2006) Differences in fecal microbiota in different European study samples in relation to age, sex, and country: a cross-sectional study. Appl Environ Microbiol 72(2):1027–1033. https://doi.org/10.1128/aem.72.2.1027-1033.2006
Munson MA, Pitt-Ford T, Chong B, Weightman A, Wade WG (2002) Molecular and cultural analysis of the microflora associated with endodontic infections. J Dent Res 81(11):761–766. https://doi.org/10.1177/0810761(Erratum in: J Dent Res. 2003 Mar; 82(3):247. J Dent Res. 2003. 82(1):69)
Muyzer G, de Waal EC, Uitterlinden AG (1993) Profiling of complex microbial samples by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59(3):695–700
Naidoo T, Konkol K, Biccard B, Dudose K, McKune AJ (2012) Elevated salivary C-reactive protein predicted by low cardio-respiratory fitness and being overweight in African children. Cardiovasc J Afr 23(9):501–506. https://doi.org/10.5830/cvja-2012-058
Nascimento GG, Leite FR, Do LG, Peres KG, Correa MB, Demarco FF et al (2015) Is weight gain associated with the incidence of periodontitis? A systematic review and meta-analysis. J Clin Periodontol 42(6):495–505. https://doi.org/10.1111/jcpe.1241767
Nascimento GG, Peres KG, Mittinty MN, Mejia GC, Silva DA, Gonzalez-Chica D et al (2017) Obesity and periodontal outcomes: a population-based cohort study in Brazil. J Periodontol 88(1):50–58
Nasidze I, Li J, Quinque D, Tang K, Stoneking M (2009) Global diversity in the human salivary microbiome. Gen Res 19:636–643. https://doi.org/10.1101/gr.084616.108
Nessner Kavamura V, Taketani RG, Lançoni MD, Andreote FD, Mendes R, Soares de Melo I (2013) Water regime influences bulk soil and Rhizosphere of Cereus jamacaru bacterial communities in the Brazilian Caatinga biome. PLoS One 8(9):e73606
Ng M, Fleming T, Robinson M, Thomson B, Graetz N, Margono C et al (2014) Global, regional, and national prevalence of overweight and obesity in children and adults during 1980–2013: a systematic analysis for the Global Burden of Disease Study. Lancet 384(9945):766–781. https://doi.org/10.1016/s0140-6736(14)60460-8
Nübel U, Engelen B, Felske A, Snaidr J, Wieshuber A, Amann RI et al (1996) Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis. J Bacteriol 178(19):5636–5643
Ozga AT, Sankaranarayanan K, Tito RY, Obregon-Tito AJ, Foster MW, Tallbull G et al (2016) Oral microbiome diversity among Cheyenne and Arapaho individuals from Oklahoma. A J Phys Anthropol 161:321–327. https://doi.org/10.1002/ajpa.23033
Palle AR, Reddy CM, Shankar BS, Gelli V, Sudhakar J, Reddy KK (2013) Association between obesity and chronic periodontitis: a cross-sectional study. J Contemp Dent Pract 14(2):168–173
Palmer RJ (2014) Composition and development of oral bacterial communities. Periodontol 2000 64:20–39. https://doi.org/10.1111/j.1600-0757.2012.00453.x
Pylro VS, Roesch LF, Ortega JM, do Amaral AM, Tótola MR, Hirsch PR, Brazilian Microbiome Project Organization Committee et al (2014) Brazilian Microbiome Project: revealing the unexplored microbial diversity—challenges and prospects. Microb Ecol 67(2):237–241. https://doi.org/10.1007/s00248-013-0302-4
Pylro VS, Morais DK, de Oliveira FS, Dos Santos FG, Lemos LN, Oliveira G et al (2016) BMPOS: a flexible and user-friendly tool sets for microbiome studies. Microb Ecol 72(2):443–447. https://doi.org/10.1007/s00248-016-0785-x
Raju SC, Lagström S, Ellonen P, de Vos WM, Eriksson JG, Weiderpass E et al (2019) Gender-specific associations between saliva microbiota and body size. Front Microbiol 10(10):767. https://doi.org/10.3389/fmicb.2019.00767
Ramette A (2007) Multivariate analyses in microbial ecology. FEMS Microbiol Ecol 62(2):142–160
Ridaura VK, Faith JJ, Rey FE, Cheng J, Duncan AE, Kau AL et al (2013) Cultured gut microbiota from twins discordant for obesity modulate adiposity and metabolic phenotypes in mice. Science 341:6150. https://doi.org/10.1126/science.1241214
Sarkar A, Stoneking M, Nandineni MR (2017) Unraveling the human salivary microbiome diversity in Indian sampless. PLoS One 12:1–17. https://doi.org/10.1371/journal.pone.0184515
Shirtcliff EA, Granger DA, Likos A (2002) Gender differences in the validity of testosterone measured in saliva by immunoassay. Horm Behav 69:62–69. https://doi.org/10.1006/hbeh.2002.1798
Sogin ML, Morrison HG, Huber JA, Mark Welch D, Huse SM, Neal PR et al (2006) Microbial diversity in the deep sea and the underexplored “rare biosphere”. Proc Natl Acad Sci USA 103(32):12115–12120
Suvan J, Petrie A, Moles DR, Nibali L, Patel K, Darbar U et al (2014) Body mass index as a predictive factor of periodontal therapy outcomes. J Dent Res 93(1):49–54. https://doi.org/10.1177/0022034513511084
Tsai F, Coyle WJ (2009) The microbiome and obesity: is obesity linked to our gut flora? Curr Gastroenterol Rep 11(4):307–313
Wang Y, Qian PY (2009) Conservative fragments in bacterial 16S rRNA genes and primer design for 16S ribosomal DNA amplicons in metagenomic studies. PLoS One 4(10):e7401
Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73(16):5261–5267
Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703
WHO Guidelines Approved by the Guidelines Review Committee Geneva (2007) World Health Organization
Wood N, Johnson RB, Streckfus CF (2003) Comparison of body composition and periodontal disease using nutritional assessment techniques: third National Health and Nutrition Examination Survey (NHANES III). J Clin Periodontol 30(4):321–327
Wu Y, Chi X, Zhang Q, Chen F, Deng X (2018) Characterization of the salivary microbiome in people with obesity. PeerJ 6:e4458. https://doi.org/10.7717/peerj.4458
Zaura E, ten Cate JM (2015) Towards understanding oral health. Caries Res 49(1):55–61. https://doi.org/10.1159/000377733
Zeigler CC, Persson GR, Wondimu B, Marcus C, Sobko T, Modéer T (2012) Microbiota in the oral subgingival biofilm is associated with obesity in adolescence. Obesity 20(1):157–164
Acknowledgements
The authors thank Embrapa environment for the sequencing (Ion Torrent) procedure support in the name of Prof. Dr. Itamar Soares de Melo and PhD. Rodrigo Gouveia Taketani.
Funding
The authors thank the funding support of São Paulo Research Foundation (FAPESP Grants #2016/02942-1, #2012/04492-2 and #2014/24804-4) and National Research Council (CNPq Grant #42344/2013-3).
Author information
Authors and Affiliations
Contributions
Conceived or designed the study: KRK, FDA, PMC, FHN; performed research: PAMA, PAG, DAS APP; analyzed data: PAMA; contributed reagents/materials/analysis tools: KRK, PMC, FDA, FHN; building and editing graphical arts: PAMA and AJS; wrote the paper: PAMA, PAG, DAS, AJS, APP, KRK, PMC, FDA, FHN.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval and consent to participate
Subjects provided written consent for enrollment and saliva collection. The Ethics Committee at the Piracicaba Dental School (protocol #152/2014) approved the study protocol and the ethical standards.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
de Andrade, P.A.M., Giovani, P.A., Araujo, D.S. et al. Shifts in the bacterial community of saliva give insights on the relationship between obesity and oral microbiota in adolescents. Arch Microbiol 202, 1085–1095 (2020). https://doi.org/10.1007/s00203-020-01817-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-020-01817-y