大学生饮食习惯与唾液微生物多样性的关联
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摘要
目的:通过16S rRNA基因高通量测序分析,研究不同饮食习惯大学生的唾液菌群多样性。方法:在大学生群体中通过问卷调查的方式,选取36位受试者,其中19位同学喜荤且饮食以荤食为主(A组),17位同学喜素且饮食以素食为主(B组)。采取唾液样本,提取DNA,聚合酶链式反应扩增,利用Illumina测序平台对16S rRNA V3～V4区进行双端测序,利用QIIME等软件进行细菌群落结构及多样性分析。结果:共获得990 286条优质序列,检测出212个可操作分类单元,归属于16个门、143个属。A组和B组唾液微生物群落多样性分析显示,在门水平上,A组中SR1、Fusobacteria、Candidatus、Saccharibacteria的丰度显著低于B组。在属水平上,A组中的优势属,如消化链球菌(Peptostreptococcus)、微单胞菌(Parvimonas)、优杆菌(Eubacterium)、Saccharibacteria_genera_incertae_sedis、SR1_genera_incertae_sedis、卡氏菌(Catonella)、艰难杆菌(Mogibacterium)、Solobacterium相对丰度均比B组显著降低(P<0.05),而A组中棒状杆菌属(Corynebacterium)相对丰度比B组显著升高(P<0.05)。结论:人类口腔唾液有复杂的微生物群落结构,本研究表明大学生人群的饮食习惯对其口腔微生物的群落结构可能有显著影响。
Objective: To study the salivary bacterial diversity of college students with different dietary habits through high throughput sequencing analysis of 16 S rRNA gene. Methods: A total of 36 subjects were selected for a questionnaire survey including 19 students with a preference for meat having a meat-dominated diet(group A), and 17 students with a preference for vegetables having a vegetable-dominated diet(group B). Saliva samples were collected to extract DNA, followed by polymerase chain reaction(PCR) amplification and high-throughput sequencing of bacterial 16 S rRNA V3–V4 region using the Illumina platform. The bacterial community structure and diversity were analyzed using the QIIME software. Results: A total of 990 286 high-quality reads were obtained and clustered into 212 operational taxonomic units(OTUs) for further analysis. A total of 16 bacterial phyla and 143 genera were detected. The diversity analysis of salivary microbiota showed that at the phylum level, SR1, Fusobacteria, Candidatus and Saccharibacteria were significantly decreased in group A as compared to group B; at the genus level, the relative abundance of the dominant genera Peptostreptococcus, Parvimonas, Eubacterium, Saccharibacteria_genera_incertae_sedis, SR1_genera_incertae_sedis, Catonella, Mogibacterium, and Solobacterium in group A were significantly lower than in group B, while the opposite was observed for the genus Corynebacterium(P<0.05). Conclusion: Human saliva has a complex microbial community structure. Dietary habits in college students(meat eaters and vegetarians)may have a significant influence on the structure of the oral microbial community.
Objective: To study the salivary bacterial diversity of college students with different dietary habits through high throughput sequencing analysis of 16 S rRNA gene. Methods: A total of 36 subjects were selected for a questionnaire survey including 19 students with a preference for meat having a meat-dominated diet(group A), and 17 students with a preference for vegetables having a vegetable-dominated diet(group B). Saliva samples were collected to extract DNA, followed by polymerase chain reaction(PCR) amplification and high-throughput sequencing of bacterial 16 S rRNA V3–V4 region using the Illumina platform. The bacterial community structure and diversity were analyzed using the QIIME software. Results: A total of 990 286 high-quality reads were obtained and clustered into 212 operational taxonomic units(OTUs) for further analysis. A total of 16 bacterial phyla and 143 genera were detected. The diversity analysis of salivary microbiota showed that at the phylum level, SR1, Fusobacteria, Candidatus and Saccharibacteria were significantly decreased in group A as compared to group B; at the genus level, the relative abundance of the dominant genera Peptostreptococcus, Parvimonas, Eubacterium, Saccharibacteria_genera_incertae_sedis, SR1_genera_incertae_sedis, Catonella, Mogibacterium, and Solobacterium in group A were significantly lower than in group B, while the opposite was observed for the genus Corynebacterium(P<0.05). Conclusion: Human saliva has a complex microbial community structure. Dietary habits in college students(meat eaters and vegetarians)may have a significant influence on the structure of the oral microbial community.
引文
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[2]TAKESHITA T,KAGEYAMA S,FURUTA M,et al.Bacterial diversity in saliva and oral health-related conditions:the Hisayama study[J].Scientific Reports,2016,6:22164.DOI:10.1038/srep22164.
[3]何金枝,徐欣,周学东.口腔微生物与全身健康研究进展[J].微生物与感染,2017,12(3):139-145.DOI:10.3969/j.issn.1673-6184.2017.03.003.
[4]AAS J A,PASTER B J,STOKES L N,et al.Defining the normal bacterial flora of the oral cavity[J].Journal of Clinical Microbiology,2005,43(11):5721-5732.DOI:10.1128/JCM.43.11.5721-5732.2005.
[5]VARTOUKIAN S R,ADAMOWSKA A,LAWLOR M,et al.In vitro cultivation of‘unculturable’oral bacteria,facilitated by community culture and media supplementation with siderophores[J].PLoS ONE,2016,11(1):e0146926.DOI:10.1371/journal.pone.0146926.
[6]MASON M R,NAGARAJA H N,CAMERLENGO T,et al.Correction:deep sequencing identifies ethnicity-specific bacterial signatures in the oral microbiome[J].PLoS ONE,2014,9(6):e99933.DOI:10.1371/journal.pone.0099933.
[7]REN Wen,XUN Zhe,WANG Zicheng,et al.Tongue coating and the salivary microbial communities vary in children with halitosis[J].Scientific Reports,2016,6:1-12.DOI:10.1038/srep24481.
[8]ATARASHI K,SUDA W,LUO C W,et al.Ectopic colonization of oral bacteria in the intestine drives TH1 cell induction and inflammation[J].Science,2017,358:359-365.DOI:10.1126/science.aan4526.
[9]CAPORASO J G,LAUBER C L,WALTERS W A,et al.Ultra-highthroughput microbial community analysis on the Illumina HiSeq and MiSeq platforms[J].ISME Journal,2012,6(8):1621-1624.DOI:10.1038/ismej.2012.8.
[10]WEINSTOCK G M.Genomic approaches to studying the human microbiota[J].Nature,2012,489:250-256.DOI:10.1038/nature11553.
[11]CLAESSON M J,WANG Q,O’SULLIVAN O,et al.Comparison of two next-generation sequencing technologies for resolving highly complex microbiota composition using tandem variable16S rRNA gene regions[J].Nucleic Acids Research,2010,38(22):e200.DOI:10.1093/nar/gkq873.
[12]WU X,ZHANG H,CHEN J,et al.Comparison of the fecal microbiota of dholes high-throughput Illumina sequencing of the V3-V4 region of the 16S rRNA gene[J].Applied Microbiology&Biotechnology,2016,100(8):3577-3586.DOI:10.1007/s00253-015-7257-y.
[13]胡欣培,李梦媛,马静,等.大学生口腔健康调查分析[J].宁夏医学杂志,2015,37(7):656-657.DOI:10.13621/j.1001-5949.2015.07.0656.
[14]廖雪梅,王雪华,熊志勇,等.胆囊结石饮食相关影响因素的横断面调查[J].中华肝脏外科手术学电子杂志,2016,5(6):398-403.DOI:10.3877/cma.j.issn.2095-3232.2016.06.013.
[15]NAVAZESH M.Methods for collecting saliva[J].Annals of the New York Academy of Sciences,1993,694(1):72-77.DOI:10.1111/j.1749-6632.1993.tb18343.x.
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[22]R Core Team.R:a language and environment for statistical computing.R Foundation for Statistical Computing[M].Vienna:Technical Report,2012:10-60.
[23]VESTY A,BISWAS K,TAYLOR M W,et al.Evaluating the impact of DNA extraction method on the representation of human oral bacterial and fungal communities[J].PLoS ONE,2017,12(1):e0169877.DOI:10.1371/journal.pone.0169877.
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[25]吴芳,李俊平,汪珍珍,等.16S rRNA基因克隆文库分析比较牙周炎患者和健康人口腔唾液微生物多样性[J].微生物学通报,2016,43(6):1295-1303.DOI:10.13344/j.microbiol.china.150792.
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[29]KILIAN M,CHAPPLE I L C,HANNIG M,et al.The oral microbiome:an update for oral healthcare professionals[J].British Dental Journal,2016,221(10):657-666.DOI:10.1038/sj.bdj.2016.865.
[30]SATO Y,YAMAGISHI J,YAMASHITA R,et al.Inter-Individual differences in the oral bacteriome are greater than intra-day fluctuations in individuals[J].PLoS ONE,2015,10(6):e0131607.DOI:10.1371/journal.pone.0131607.