Media supplements and methods to culture human gastrointestinal anaerobic microorganisms

Abstract

A media supplement for culturing anaerobic bacteria is provided which comprises a filtrate of effluent from a chemostat vessel in which a target bacterial ecosystem has been cultured. Methods of using the supplement for culturing or isolating anaerobic microbial strains or communities, particularly anaerobic bacteria from the human gut, are also provided.

Description

RELATED APPLICATIONS[0001]This application claims priority to U.S. provisional application No. 61 / 534,456, filed on Sep. 14, 2011, the entire contents of which are hereby incorporated by reference. This application is related to copending international PCT application titled “Method for Treatment of Disorders of the Gastrointestinal System,” filed Sep. 14, 2012, the entire contents of which are hereby incorporated by reference.FIELD OF THE INVENTION[0002]This invention relates to methods for culturing microbial communities from the human distal colon and methods for isolating anaerobic microbes from such communities, as well as media supplements for use in such methods.BACKGROUND OF THE INVENTION[0003]The human gut is the most densely inhabited ecosystem on Earth (Marchesi and Shanahan, 2007). Like other complex microbial ecosystems, the human microbiota has not been sampled to completion (Eckburg et al., 2005). This is because the individual species of the gut microbiota are diffic...

AI Technical Summary

Benefits of technology

The patent text describes methods for culturing microorganisms from the human large intestine and for developing microbial communities from the human distal colon. The methods involve using a media supplement called "Liquid Gold," which is a filtrate of effluent from a chemostat vessel in which a target bacterial ecosystem has been cultured. The media supplement can be used to culture anaerobic bacteria that are difficult to isolate using traditional methods. The methods can also involve using a single-stage chemostat and collecting effluent from the chemostat to produce the media supplement. The patent text also describes the use of Liquid Gold in the development of microbial communities from human fecal samples. Overall, the patent text provides methods for culturing and characterizing microorganisms from the human large intestine and for developing microbial communities from the human distal colon.

Problems solved by technology

This is because the individual species of the gut microbiota are difficult to culture axenically in vitro (Hart et al., 2002).

It is recognized that novel culture techniques are required to grow these “unculturable” microorganisms.

Studies of gut microbiota have been hampered by a lack of model systems.

For example, different study participants can each have unique, host-specific community profiles representing their gut microbiota, making comparison of the gut microbiota between subjects difficult, especially when attempting to correlate the effects of a treatment to changes in the gut microbiota.

In vivo models also often limit the dynamic monitoring of the gut microbiota by deriving their data from end-point measurements.

Experiments involving humans or animals require research ethics approval which can limit the experiments conducted on an individual's gut microbiota in vivo.

However, while chemostats provide a useful tool to investigate the microbial ecology of the gut, operational parameters vary widely between different models in different laboratories, often without experimental validation.

Currently, there is a lack of standardization between DGGE analysis methods used in different research laboratories.

However, laboratories utilizing these analysis programs do not use consistent methods when analyzing their DGGE gels and report varying data on their communities.

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