McBain A J, Sissons C, Ledder R G, Sreenivasan P K, De Vizio W, Gilbert P
School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Manchester, UK.
J Appl Microbiol. 2005;98(3):624-34. doi: 10.1111/j.1365-2672.2004.02483.x.
To validate perfused, inline, filter-based fermentation systems (multiple Sorbarod devices, MSD) for their ability to maintain stable oral bacterial communities. MSD enable replicate (n=5) microcosm biofilms (BF) to be established and sampled, together with their perfusates (PA, cells in eluted medium).
Fresh saliva from human volunteers was used to inoculate MSD, incubated in an anaerobic cabinet and perfused with artificial saliva at 7 ml h(-1). BF within Sorbarod filters and cells eluted in the PA were analysed at 24-h intervals by differential bacteriological culture and checkerboard DNA-DNA hybridization (CKB, 40 oral species). Dynamic stability was apparent after 2-3 days within both BF and PA as evidenced by culture, CKB data and pH measurements. BF harboured large numbers of anaerobic species and facultative anaerobes [ca 10-11 log10 colony-forming units (CFU)/filter] comprising considerable numbers of streptococci and Gram-negative species. PA contained ca 9-10 log(10) CFU ml(-1) suggesting an apparent mean growth rate of 0.1 h(-1) for the BF, as a whole corresponding to a mean generation time of 10 h. CKB analysis revealed considerable bacterial diversity within the respective MSD. Inter-individual variations in the relative species abundance of inocula was broadly reproduced in the MSD (BF and PA), although considerable variation was apparent between triplicate models established using saliva from one saliva donor or from three individual donors. The dominance of Gram-negative species, indicated by culture was supported by CKB analysis (major species, Prevotella melaninogenica and Fusobacterium nucleatum).
Data obtained from the various analytical approaches showed a high degree of congruence. The MSD enables the maintenance of complex, stable salivary microcosms and represents a simple, reproducible tool for modelling individual oral bacterial ecosystems.
This study demonstrates the utility of the MSD for studying the micro-ecology of the oral cavity.
验证灌注式、在线、基于过滤器的发酵系统（多个索巴罗德装置，MSD）维持稳定口腔细菌群落的能力。MSD能够建立并取样重复的（n = 5）微观生物膜（BF）及其灌注液（PA，洗脱培养基中的细胞）。
使用人类志愿者的新鲜唾液接种MSD，置于厌氧箱中培养，并用人工唾液以7 ml h⁻¹的流速进行灌注。每隔24小时通过差异细菌培养和棋盘式DNA-DNA杂交（CKB，40种口腔菌种）分析索巴罗德过滤器内的BF以及PA中洗脱的细胞。培养、CKB数据和pH测量结果表明，BF和PA在2 - 3天后呈现出动态稳定性。BF中含有大量厌氧菌和兼性厌氧菌[约10 - 11 log₁₀集落形成单位（CFU）/过滤器]，包括大量链球菌和革兰氏阴性菌。PA中含有约9 - 10 log₁₀ CFU/ml，表明BF的表观平均生长速率为0.1 h⁻¹，总体平均世代时间为10小时。CKB分析显示各个MSD内细菌多样性显著。接种物相对菌种丰度的个体间差异在MSD（BF和PA）中大致重现，尽管使用一名唾液供体或三名个体供体的唾液建立的一式三份模型之间存在明显差异。培养显示革兰氏阴性菌占优势，CKB分析（主要菌种，产黑色素普雷沃菌和具核梭杆菌）也支持了这一结果。
从各种分析方法获得的数据显示出高度一致性。MSD能够维持复杂、稳定的唾液微观世界，是模拟个体口腔细菌生态系统的简单、可重复工具。
本研究证明了MSD在研究口腔微生态方面的实用性。
