Ledder R G, Gilbert P, Pluen A, Sreenivasan P K, De Vizio W, McBain A J
School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Manchester, UK.
J Appl Microbiol. 2006 May;100(5):1123-31. doi: 10.1111/j.1365-2672.2006.02847.x.
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).
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.
Data indicate that marked inter-individual variations in human salivary composition can be partially replicated in individualized MSD microcosms.
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.
研究多重索巴罗德装置（MSD）在再现个体间口腔微生物群差异方面的功效。MSD可支持在平行纤维素滤膜上的固着生长，滤膜用人工唾液灌注。这使得生物膜（BF）得以生长和采样，同时还能采集洗脱液（灌注液，PAs）中释放的细胞。
建立了两组一式三份的MSD。一组用来自三名不同志愿者的新鲜唾液接种；第二组从一名唾液捐赠者处接种。两组均在厌氧箱中孵育。通过对16S rDNA进行PCR-变性梯度凝胶电泳（DGGE），每隔24小时对BF和PA进行分析。构建层次聚类树状图，以便根据群落相关性随时间对群落指纹进行分类。DGGE（>20条不同的DGGE条带）和共聚焦扫描激光显微镜表明，MSD支持复杂的口腔群落。DGGE条带测序揭示了细菌多样性以及厌氧物种的高发生率，包括普氏菌属。树状图显示，来自不同志愿者（DV）的唾液接种物以及每个相关MSD中相对物种丰度存在明显的个体间差异（均>45%，大多数约85%一致）。使用来自单一志愿者（SV）的唾液建立的一式三份模型之间的差异较小（均>58%；大多数约95%一致）。在大多数DV MSD病例中，PAs与相关生物膜和接种物聚集在一起，而SV MSD群落图谱则聚集在重复的MSD之间。
数据表明，人类唾液成分中明显的个体间差异可在个体化的MSD微观世界中部分重现。
本研究证明了个体口腔微生物群的体外再现，并表明考虑个体间变异性将提高微观世界研究的相关性。
