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使用微型生物反应器发酵系统作为远端结肠的可重复、高通量批次模型。

The Use of a Mini-Bioreactor Fermentation System as a Reproducible, High-Throughput Batch Model of the Distal Colon.

作者信息

O'Donnell Michelle M, Rea Mary C, Shanahan Fergus, Ross R P

机构信息

APC Microbiome Institute, University College Cork, National University of Ireland, Cork, Ireland.

Teagasc Food Research Centre, Moorepark, Fermoy, Cork, Ireland.

出版信息

Front Microbiol. 2018 Aug 10;9:1844. doi: 10.3389/fmicb.2018.01844. eCollection 2018.

Abstract

colon fermentation systems are highly versatile as models for analyzing gastrointestinal tract microbiota composition and functionality. colon models range in size and functionality from bench-top micro fermenters to large units housed in individualized cabinets. The length of set-up time (including stabilization periods) for each fermentation system can range from hours to weeks to months. The aim of this study was to investigate a single-use cassette mini-fermentation system as a reproducible batch model of the colon. The online data log from the cassettes (triplicate wells across four different cassettes, = 12) was sensitive enough to identify real-time changes in pH, temperature, dissolved oxygen or liquid addition (sodium hydroxide) during the runs which could be addressed if an alarm set-point was triggered. The alpha diversity indices also showed little variation between cassettes with the samples clustering around the mean. The weighted beta diversity PCoA analysis illustrated that 95% of the variance between the samples was accounted for by the time-point and not the fermentation run/cassette used. The variation in taxonomic diversity between cassettes was limited to less than 20 out of 115 genera. This study provides evidence that micro-bioreactors provide some very attractive advantages as batch models for the human colon. We show for the first time the use of the micro-Matrix a 24-well sophisticated parallel controlled cassette-based bioreactors as a batch colon model. We demonstrated a high level of reproducibility across fermentation cassettes when used in conjunction with a standardized fecal microbiota. The machine can operate 24 individual fermentations simultaneously and are relatively cost effective. Based on next generation sequencing analysis, the micro-bioreactors offer a high degree of reproducibility together with high-throughput capacity. This makes it a potential system for large screening projects that can then be scaled up to large fermenters or human/animal experiments.

摘要

结肠发酵系统作为分析胃肠道微生物群组成和功能的模型具有高度的通用性。结肠模型在大小和功能上各不相同,从台式微型发酵罐到放置在独立机柜中的大型装置。每个发酵系统的设置时间(包括稳定期)可能从数小时到数周再到数月不等。本研究的目的是研究一种一次性盒式微型发酵系统,作为结肠的可重复批次模型。来自盒式装置(四个不同盒式装置中的三个重复孔,共12个)的在线数据记录足够灵敏,能够识别运行过程中pH值、温度、溶解氧或液体添加(氢氧化钠)的实时变化,如果触发警报设定点,这些变化可以得到处理。α多样性指数在不同盒式装置之间也显示出很小的差异,样本聚集在平均值周围。加权β多样性主坐标分析表明,样本之间95%的差异是由时间点造成的,而不是所用的发酵运行/盒式装置。不同盒式装置之间分类多样性的差异仅限于115个属中的不到20个。本研究提供了证据,表明微型生物反应器作为人类结肠的批次模型具有一些非常吸引人的优势。我们首次展示了使用微型基质,一种基于24孔复杂平行控制盒式的生物反应器作为批次结肠模型。当与标准化的粪便微生物群一起使用时,我们证明了不同发酵盒式装置之间具有高度的可重复性。该机器可以同时运行24个独立发酵,且成本相对较低。基于下一代测序分析,微型生物反应器具有高度的可重复性和高通量能力。这使其成为大型筛选项目的潜在系统,然后可以扩大规模用于大型发酵罐或人类/动物实验。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddf2/6096000/5a1c53886e82/fmicb-09-01844-g001.jpg

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