Individual microflora beget unique oral microcosms
Individual microflora beget unique oral microcosms
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DOI：
10.1111/j.1365-2672.2006.02847.x
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发表时间：
2006-05-01
影响因子：
4
通讯作者：
McBain, A. J.
中科院分区：
文献类型：
--
作者：
Ledder, R. G.;Gilbert, P.;McBain, A. J.
目的：检查多Sorbarod装置（MSD）再现口腔微生物菌群个体间差异的有效性。MSD支持平行纤维素过滤器上的固着生长，用人工唾液灌注。这使得生物膜（BF）能够与洗脱培养基（灌注液，PA）中释放的细胞一起生长和取样。使用来自三个独立志愿者的新鲜唾液接种;第二组从一个唾液供体接种。两者均在厌氧箱中孵育。BF和PA进行了分析，在24小时的时间间隔，通过PCR-变性梯度凝胶电泳（DGGE）的16 S rDNA。构建了层次树状图，以便根据社区相关性对社区指纹进行排序。MSD支持复杂的口腔群落，如DGGE（> 20个不同的DGGE条带）和共聚焦扫描激光显微镜所证明的。DGGE条带测序揭示了细菌学多样性和厌氧物种的高发生率，包括普雷沃氏菌属。树状图显示了来自不同志愿者（DV）和每个相关MSD的唾液接种物内的相对物种丰度的显著个体间差异（所有> 45%，大多数c. 85%的一致性）。在使用来自单个志愿者（SV）的唾液建立的一式三份模型之间显示较小的变化（全部> 58%;大多数c. 95%一致性）。在大多数情况下，对于DV MSD，PA与相关的生物膜和接种物聚集在一起，而SV MSD群落特征在重复的MSD之间聚集。结论：数据表明，人类唾液组成的显著个体间差异可以在个体化的MSD微观世界中部分复制。这项研究证明了个体口腔微生物的体外繁殖，并表明考虑个体间的变异性将增加微观世界研究的相关性。
Aims: To examine the efficacy of the multiple Sorbarod device (MSD) for the reproduction of inter-individual variations in oral microbiotas. The MSD supports sessile growth on parallel cellulose filters, perfused with artificial saliva. This enables biofilms (BF) to be grown and sampled, together with released cells in eluted medium (perfusates, PAs).Methods and Results: Two sets of triplicate MSDs were established One set. was inoculated using fresh saliva from three separate volunteers; the second set was inoculated from one saliva donor. Both were incubated in an anaerobic cabinet. BF and PA were analysed at 24-h intervals by PCR-denaturing gradient gel electrophoresis (DGGE) of 16S rDNA. Hierarchical dendrograms were constructed in order to sort community fingerprints over time, based on community relatedness. The MSD supported complex oral communities, as evidenced by DGGE (> 20 distinct DGGE bands) and confocal scanning laser microscopy. DGGE band sequencing revealed bacteriological diversity and a high incidence of anaerobic species, including Prevotella sp. Dendrograms demonstrated marked inter-individual variation in the relative species abundance within salivary inocula from different volunteers (DV) and each associated MSD (all > 45%, majority c. 85% concordance). Less variation was shown between triplicate models established using saliva from a single volunteer (SV) (all > 58%; majority c. 95% concordance). PAs clustered together with the associated biofilms and inocula in the majority of cases for the DV MSDs whilst SV MSD community profiles clustered between replicate MSDs.Conclusions: Data indicate that marked inter-individual variations in human salivary composition can be partially replicated in individualized MSD microcosms.Significance and Impact of the Study: This study demonstrates the in vitro reproduction of individual oral microbiotas and suggests that taking inter-individual variability into account will increase the relevance of microcosm studies.