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Culturing of ‘unculturable’ human microbiota reveals novel taxa and extensive sporulation

alicec

Senior Member
Messages
1,572
Location
Australia
http://www.nature.com/nature/journal/vaop/ncurrent/full/nature17645.html

Culturing of ‘unculturable’ human microbiota reveals novel taxa and extensive sporulation
Hilary P. Browne,1, * Samuel C. Forster,1, 2, 3, * Blessing O. Anonye,1, Nitin Kumar,1, B. Anne Neville,1, Mark D. Stares,1, David Goulding4, & Trevor D. Lawley1,
Journal name Nature Year published: (2016)
DOI: doi:10.1038/nature17645 Received 25 September 2015 Accepted 08 March 2016
Published online 04 May 2016

Our intestinal microbiota harbours a diverse bacterial community required for our health, sustenance and wellbeing1, 2. Intestinal colonization begins at birth and climaxes with the acquisition of two dominant groups of strict anaerobic bacteria belonging to the Firmicutes and Bacteroidetes phyla2. Culture-independent, genomic approaches have transformed our understanding of the role of the human microbiome in health and many diseases1. However, owing to the prevailing perception that our indigenous bacteria are largely recalcitrant to culture, many of their functions and phenotypes remain unknown3. Here we describe a novel workflow based on targeted phenotypic culturing linked to large-scale whole-genome sequencing, phylogenetic analysis and computational modelling that demonstrates that a substantial proportion of the intestinal bacteria are culturable. Applying this approach to healthy individuals, we isolated 137 bacterial species from characterized and candidate novel families, genera and species that were archived as pure cultures. Whole-genome and metagenomic sequencing, combined with computational and phenotypic analysis, suggests that at least 50–60% of the bacterial genera from the intestinal microbiota of a healthy individual produce resilient spores, specialized for host-to-host transmission. Our approach unlocks the human intestinal microbiota for phenotypic analysis and reveals how a marked proportion of oxygen-sensitive intestinal bacteria can be transmitted between individuals, affecting microbiota heritability.
 

alicec

Senior Member
Messages
1,572
Location
Australia
I found this yesterday while looking for something else. Then coincidentally last night I heard one of the authors interviewed on an Australian radio program.

I got quite excited reading the paper and this was only reinforced when I heard Sam Forster say they have all these spores stored in their freezer and within a few years he could see them becoming the basis for new generation probiotics that would replace FMT in recolonising damaged guts (well that is my paraphrase, I can't remember exactly what he said).

This work is the type of breakthrough we need to translate the explosion of new information on the gut microbiota derived from DNA sequencing into treatments that can make a real difference.

There are two things of significance.

First they have found ways of culturing many of these previously unculturable anaerobes. This means they can be thoroughly studied and their function better understood.

Even more importantly, in finding that many of them can be induced to form spores, they have found a way around the limitations of current probiotics.

Apart from the fact that current probiotic species don't in any way reflect the species actually in the gut, they do not colonise it either. Their benefit appears to derive almost entirely from modulation of the host immune system as they transit through.

Spores are a way in which bacteria survive removal from a beneficial environment, a kind of dormant state, which enables the organism to survive for an enormously long time until it again encounters a suitable environment for growth.

So now we have stable forms of many of the dominant gut bacteria (plus others we know little about) which could form the basis for future probiotics , along with ways of studying them so we can determine which are the critical core that might be able to repair damaged guts.

This could well lead to the kind of tailored probiotics that we really need, rather than the hit and miss alternative of FMT.
 

Richard7

Senior Member
Messages
772
Location
Australia
Scientifically this should be fascinating, but I wonder about making these things into pills.

I recently read this interview with Jorge Frias-Lopez where he explains that what matters when looking at the ecosystem of a dental plaque is the genes that the bacteria have the potential to express. Because when the environment changes bacteria that had being commensal or beneficial can change in to pathogens if they have the genes to do so. http://www.nidcr.nih.gov/ScienceSpo...ral-microbiome-genomics-and-periodontitis.htm

This is obviously a potential weakness in FMT.

FMT's strength, when compared to normal probiotic supplementation is that they disrupt the existing ecosystem by washing most of it away and then implant a whole healthy ecosystem, and repeat this several times.

When you adding or a couple of dozen species by taking a pill they are much less likely to find ecological niches. They are much less likely to survive in the gut.

I would think that they are also much more likely to behave unexpectedly when you are throwing them into an entirely new environment. I would expect that they would be much more likely to either switch on pathogenic genes or to cause existing bacteria to do the same.

This could also happen with FMT, each person is a new environment. Each diet is a new environment. Maybe There would be a problem, but at least we know that we have a gang of microbes that can play nice. And have presumably upregulated and down regulated the genes necessary for this to be so.

I don't know that this would be the case in a probiotic pill.

If this was very carefully researched before being made into a pill, with every microbe being tested for potentially pathogenic genes, it could be great. But I kind of expect that we will get a lot of shoddy supplements that are over hyped and could be dangerous.

Sort of like all of medicine there I guess.
 

Richard7

Senior Member
Messages
772
Location
Australia
Oh and I should add i have used, and continue to use probiotics. I am using oral ones at the moment.

But I have reacted badly to some of them and have been glad that they were not spore forming and unlikely to last long in my gut.
 

alicec

Senior Member
Messages
1,572
Location
Australia
When you adding or a couple of dozen species by taking a pill they are much less likely to find ecological niches. They are much less likely to survive in the gut.

I would think that they are also much more likely to behave unexpectedly when you are throwing them into an entirely new environment. I would expect that they would be much more likely to either switch on pathogenic genes or to cause existing bacteria to do the same.

That is all just assumption. We don't know because we have never been able to study it.

If this was very carefully researched before being made into a pill, with every microbe being tested for potentially pathogenic genes, it could be great.

The study has developed at least some of the tools to enable us to do just that.

But I kind of expect that we will get a lot of shoddy supplements that are over hyped and could be dangerous.

I think we will be well able to pick out the charlatans. There are also plenty of dedicated scientists and clinicians out there who truly want to come up with better solutions. I for one am very willing to urge them on.
 

Richard7

Senior Member
Messages
772
Location
Australia
Alicec, I want to see the science done too. I just have these moments, like when I read the interview I linked to above where I get a sense of how much there is to know before we can be sure that we are intervening safely.

And how vastly oversimplified and misleading much of the information I have read (or heard from my doctors) has been and how dodgy the interventions I have taken have been.

re this point
When you adding or a couple of dozen species by taking a pill they are much less likely to find ecological niches. They are much less likely to survive in the gut.

I would think that they are also much more likely to behave unexpectedly when you are throwing them into an entirely new environment. I would expect that they would be much more likely to either switch on pathogenic genes or to cause existing bacteria to do the same.

That is all just assumption. We don't know because we have never been able to study it.

I understand the first half of my statement to be true. I am not an ecologist and have no training in it. But my understanding is that when a new animal or plant is introduced to an intact ecosystem the general rule is that it won't find a niche and become established.

There are exceptions, in New Zealand there were no plants that were good at colonising recently disturbed ground until blackberries were introduced. But I have only heard of it because I listened to a talk on the radio where an ecologist presented it as an exception.

I have read the case of the rabbit in Australia being presented as more usual. There were two or three unsuccessful attempts to introduce it into a reasonably intact ecosystem and then when it was introduced to a massively degraded ecosystem there was an overly successful one.

But have heard more than one ecologist state that a handful of bears or some other sort of animal being let loose in a foreign ecosystem will, even if they survive for a generation or two, most likely soon die out.

I have read of practitioners having issues trying to introduce beneficial bacteria to their patients guts, and have myself being such a patient.

So I do indeed assume that this is the case. Am I wrong? I mean wrong about the theory, no one knows about the practice as there is none yet.



For the second part, sure I do not know what the case is. We won't know until the experiment is done. That is kind of why I am expressing doubts rather than making statements.

But in that article that I linked to there is a section before the interview which starts with a quote from Philip Marsh

“It is a basic tenet of microbial ecology that a major change to an ecosystem produces a corresponding disturbance to the resident community,” he explained. “Examples of this have been reported in ecosystems as diverse as soil, skin, and water.”

As Marsh’s hypothesis gained adherents in the 1990s, periodontal research adapted a number of existing ecological terms. Most notably, dental plaque became a biofilm, a dynamic, polymicrobial community that, like lichens growing along a craggy coast, colonizes various and sundry pits, fissures, and other oral surfaces. From this ecological perspective, the way forward in periodontal research involved gaining a more detailed understanding of these microbial ecosystems, the environmental disturbances that destabilize them, and the opportunistic pathogens that exploit the imbalance to cause disease.

And from the interview with Jorge Frias-Lopez
What was most interesting about the new organisms?

There were a lot of organisms that are not considered at this point to be periodontal pathogens that were producing large numbers of putative virulence factors. I say, “putative” because this is based on homology.

Meaning the gene sequences, to various degrees, resemble known virulence factors?

Right. It may turn out that not all of them are virulence factors. But the huge number of them being produced suggests to me that the whole community is becoming more virulent, not just the Red Complex organisms. Somehow the new environment induces the production of these virulence factors in the whole community. It’s just a competition to make them. To me, that was the most surprising and interesting. It indicated that the community becomes more dysbiotic and more virulent under those environmental conditions.

Obviously they are speaking of disturbances in the ecosystem initiated by a pathogen.

Which is kind of what we are talking about with FMT. I do not know the details, but I understand that for FMT they use some medical intervention or other to stop inflammation in the gut, wash it out then apply the sample of a healthy microbiome.

Not asserting, just hypothesising, but would this not mean from the microbes points of view they were going from a healthy gut to a seemingly healthy gut. So they would be less likely, I once again hypothesise, to start producing those virulence factors or putative virulence factors mentioned by Frias-Lopez.

I was kind assuming that the pill would be assumedly commensal or beneficial bacteria dropped into a diseased gut. That it would be just like the probiotics I have taken. And that if we did not fully understand them, which seems to be the case with stuff on the market now, and the assumption that they were certainly commensal or beneficial proved to be wrong etc etc etc.

I do not mean to be all doom and gloom. One could presumably do the science and do it properly and all would be great. It is just that that article made an impression on me, and I have being thinking of some of the risks I took without knowing them to be risks when I listened to doctors and microbiologists and biochemists who were very certain that they knew what they were doing.

I don't mean to be doom and gloom but maybe I'm having one of those days. Perhaps all these points are irrelevant and I should say I cannot wait till the research is done, and someone has written a book or an article about it that is accessible and not too simplified or too wrong. That too is an assumption, of course, and one I assume I would like to put to the test.