[Music] yeah I think measuring degrading enzymes would be a good way of of seeing how efficient the microbiota is yes so I would say is the definite yes for the first question because of course you need the enzymes there to degrade but as I showed you also it's often the whole three-dimensional structure that is important but of course we're doing that essentially if we incubating our microbiota or specific bacteria with these recalcitrant substrates and you see it break down and the substrate is a more direct way well the se of a profile is much more simple than the microbiota profile so I think yeah I think if you the SEO of a profile tells you something about whether person's likely to have a healthy microbiota and a healthy meta but metabolite profile but it you can't really go back from a very simple profile to the complexity in the microbiota so I think it wouldn't be a good indicator for microbiota I would go for molecular techniques for that I think the other questions about the individual genetic variants for human but for animal production uniform production so one important issue what's your perspective for for that well that's a really I think the the interplay between the human genetics and the microbiota composition isn't really fully understood yet so I don't think I can give a clear answer to this there are definitely connections some people associate even different blood groups with different types of microbiota it's also not really clear how individual specific the colonization is on the other hand we have all the fecal transplantation work now and it does look like that some people respond very well to fecal transplantation this is more than currently to eradicate Presidium difficile infections but I think in some people the new microbiota tests which is quite well in some people there's a mix between you and the old so I think it's too early to really say how much that hosts genetics influence this the other question I rather to see like Ken that from Joran can this be achieved to accent in his enzymes as well I think this question also relates you you know nah for plants fibroids were complicated the structure particularly in a crystalline structure that's actually the bottleneck so from the animal protection for utilization of those polysaccharide so I think this question is also important yesterday okay so it's five are degrading enzymes you mean yeah so I think there's a there is a pro and con there as well of course if you judge it if you degrade them down to their monosaccharides there will not be a food source for the microbes anymore because then the animal will take most of that in so from a microbiota and short fatty acid producing point of view i don't think would be good to completely degrade the fiber but to make it maybe more accessible may be a good thing but on the other hand at least from a human perspective i think you don't want this really totally soluble rapidly fermentable fiber either because then you get a huge activity in the proximal colon people may get gut symptoms but also then very quickly runs out because it's really it's a very buffet for everybody so the bacteria can all bacteria can chip in on most bacteria can ship in right away huge activity in the proximal colon and then very little happening in the distal coolin so I think you want a mix of hard digestible stuff that genuinely takes longer to digest and you need a longer chain of bacteria to nibble away and slowly trickle it into the system that you have fermentation still further down and that probably would help to minimize the risk of that diseases further down I think I would I think I would still have some time for questions like coffee start the 11 o'clock correct yeah yeah if we can Republican go through some questions again like I think you know like I in human the main intervention are even adult to boost endogenous protection of a Pyrrhic know you what is in a passed away to go into and some without conversation okay what's the best diet to have hi beat reproduction I guess that's how I interpret the question I think it is individual to a degree but you remember my in vitro data which does show that certain fibers are more stimulating to overall Petri production or overall protein a production so I think fructose are a good source for butyrate at least in the human system and starches are also good room in a caucus bra me I by the way where I showed you as the main keystone species for starch degradation does not produce betrayed itself so the butyrate production is not by direct production from rumen Cox brew Mia this is cross feeding effects but the rumen akagi's bro Mia actually grows quite poorly in the lobby also having trouble to grow it it needs a lot of vitamins so it also is normally not there and very high amounts in people I think we boosted it so much in our human study because we gave the system so much starch so probably even in the gut it's it's actually adapted to be a slow grower it doesn't take all that much of the starch for itself I believe or I assume a lot of the starch it makes available to other bacteria and then you have future I producing bacteria that can then turn the breakdown products of the recalcitrant starch starch into butyrate but to a degree it may again be an individual difference in different people's so I think we could design diets that will generate high bit rate in most people but we need to be aware that in some people it could be different this has also been shown for lactate many years ago where there were in vitro incubations with lactate and I've shown you that some bacteria turn lactate into butyrate and others into propionate and I think there were three people in the study - were very beautiful Jenny and one person almost converted all the locked Aten to propionate so there definitely can be differences between different people it's not necessarily a one-size-fits-all I think every other question is a pretty simple question do you think a different species have similar microbiota that shouldn't be pretty good question to answer yeah I think mostly we would say if you find the same species in different animals they tend to be quite similar so even I showed you the cellulose degraders which are mainly present in the rumen because they're cellulose degradation the plant cell wall cellulose is so important for feeding the rumen animal itself so this bacterium room in the caucus Flavia fashions has this cellulose oum this enzyme apparatus to degrade cellulose and the room in the caucus sample analysis I showed you from the human gut is a close cousin it's a very related species in the same genus and it seems to have a very similar rule in the human gut so yes I think there again it's biology its nature it's messy I can't really give a blank that they're all going to be the same but mostly we find the same species in different animals has quite similar functions so next question is the activity of the starch and fiber degrading and that would be good indicator for them good microbiota yes I would say so yeah I think there is a link between pH and health so again I'm not an expert on the physiological side of things but I think the the lowering of the pH is stimulating betray producers so I think what what we find in vitro in our fermenter studies and also in our batch cultures if the pH is around over six the Bacteroides seem to be more fit the Bacteroides is winning out the beat reproduce the firm acute is hanging in there but they're they're not growing as fast so I think that the pH goes down really means the Firmicutes have more of a chance and don't get me wrong it's not all Bacteroides this and all the firm acuter so that was in the firm acutest we have some that are more page tolerant and less you may remember one of my slides with the two rows boria's which were different but i think if you want good wheat rye production lowering of the pH is important otherwise the butyrate producers will just not be able to compete that well to the bacterial deities which are propionate producers but on the other hand you can spiral out of control in ruminants there's something known as lactic acidosis which you may be aware of if the system spirals out of control and the pH drops below a certain point if it gets sort of below into the five below five region then the whole system can spiral out of control because most bacteria can't grow anymore and if you are taking over now so even in ilustrate of colitis colitis patients do some find some patients and severe colitis which have a lot of pay for the bacteria and of course everybody thinks before the bacteria of really has promoting and really wonderful but actually if they're taking over and suddenly there's only bifida bacteria it's not good anymore because they're locked a producers and if the beauty producers can't go for it anymore then that's not a good thing and they're probably then assigned for the system has gone out of control and the bacteria are just more Hardy they can withstand the low pH better so yes pH lowering is important but not too much so again if you give too much substrate for the bacteria and one go there's a higher risk of this going out of control a couple more questions I think a two of them had had asked heads answered already similarly you know and I think a lot last question would be Quito doubt so do you expect if we are on keto doubt Oh 10:00 a.m. you to fall down and how do you propose continue on the keto diet just don't do it I think it's just it's just no need they are better ways to lose weight and be healthy I just I just don't even want to think of a way to now counteract the bad effects of a keto diet it's just a bad idea I'll take em off take em up you are a builder it can reduce the pH protection is important for many things it's not just the charge in fatty acids fiber is also important for water holding and transit time and yes they ask you simple questions you mentioned the phytochemicals also affect the microbiota I just wondering in what kind of way approach very complex so yeah you can see there's many different ways some phytochemicals may inhibit certain bacteria that can be toxic to a bacteria so you potentially changed the profile in the microbiota of course people want to believe that phytochemicals are bad for proteobacteria which we don't want to have so much of but leaving other bacteria alone it's not going to be that simple but yeah you can have an inhibitory effect but more importantly the microbiota is very closely involved in the bio transformation of many phytochemicals so a lot of the phytochemicals are covalently linked to the fiber for example as I showed you and other phytochemicals are soluble in the vacuole of the plant but they're linked to sugars so if they're glycosylated then basically we can take them up and then also only even as glucose we have some enzymes that can cleave these sugars off but a lot of them cannot be digested by ourselves we need the microbiota to liberate these phytochemicals the egg-like owns the actual polyphenols to either then have their action or to be absorbed a lot of them can only be absorbed once they have been d glycosylated by the microbiota and then even the aglycone is further transformed the rocks elated D hydrogenated or hydrogenated by different bacteria and then their bio activity may change so some of these phytochemicals only have their health beneficial action once they have been further metabolized by the microbiota okay my second technical question you talked about that you meet your sub culturing because you don't do no modeling they are usually in which you're culturing what kind of a medium do you use them because of that very critical yeah the selection of a medium will smack the result and so conclusion yes it's very crucial and it's a there is no one medium that is perfect for all but the raw it really comes from a room and background so when we moved into human research about twenty years ago we used the media that were used for rumen bacterial growth so we use a lot this medium mm2 which has room and fluid in it okay and if you compare any human fecal incubation if you compare this medium over bhi which is used by many Braden heart infusion you get roughly ten times the number of bacteria so it's much better but four out unknown work for example with the vitamin we had to go to the other extreme yet we had to take everything out there could be no yeast extract not even norm come as amino acids and it's getting very hard to grow these some of these bacteria in that so okay thank you so much and Patrick got lots of questions [Applause]