448 replies to this topic

[ekaitz]

I read that the optimum temperature for kefir grains is 22C, whereas Reuteri grows best at 37C. So, not preferable.

 

Does yogurth making takes place at that temperature, 37º?

[TerryFirmer]

Does yogurth making takes place at that temperature, 37º?

 

Yes.

[Ovidus]

So correct me if I'm wrong but for easily making a high bacterial count probiotic "yogurt" with the BioGaia. The best would be a lactose free milk with glucose added so both strains multiply closer to the same rate? Has anyone figured out yet how to go the next step and encourage the strain we want to grow faster? Also am I correct in assuming a light sprinkling of some b complex from a capsule would also be a good idea for their growth rate?

 

I am curious about one thing: 

Lactose-free milk has simply been treated enzymatically and the lactose in it has been broken down into galactose and glucose, right?

When we say that using lactose-free milk will make a difference, we  assume that the L.Reuteri 6475 is not capable of breaking down lactose into its constituents. Is that really so?
Can you guys share some data?

Also -if one were completely unconcerned with taste- would it be even better to just try to "ferment" the crushed tablets in glucose-water?

 

And, acacia fibre was mentioned. 1) Any point in adding that to the yogurt during fermentation? 2) If that is not exactly a good idea, what is the ideal time of intake for the acacia fibre? Right around the consumption of Reuteri yogurt or away from it?

 

Finally, can you guys share the amount of glucose added and the exact yogurt making methods?

 

Many thanks to all

[granmasutensil]

I am curious about one thing: 

Lactose-free milk has simply been treated enzymatically and the lactose in it has been broken down into galactose and glucose, right?

When we say that using lactose-free milk will make a difference, we  assume that the L.Reuteri 6475 is not capable of breaking down lactose into its constituents. Is that really so?
Can you guys share some data?

Also -if one were completely unconcerned with taste- would it be even better to just try to "ferment" the crushed tablets in glucose-water?

 

And, acacia fibre was mentioned. 1) Any point in adding that to the yogurt during fermentation? 2) If that is not exactly a good idea, what is the ideal time of intake for the acacia fibre? Right around the consumption of Reuteri yogurt or away from it?

 

Finally, can you guys share the amount of glucose added and the exact yogurt making methods?

 

Many thanks to all

 

Hi Ovidus I don't have any scientific knowledge on the subject that's why I made my previous post to confirm what seemed to be the facts so far for making a yogurt with the probiotic.

 

But I can touch on your question about the lactose. No clue if that's how they removed the lactose in general for these lactose free milks but assuming you are right and that could be an issue I just wanted to respond and share what I was planning on using from the start.

 

In the USA you can buy lactose free milk products from a brand called fairlife, https://fairlife.com/our-process/ . I originally planned to choose this product/brand of milk to make the yogurt because someone else in this thread said it's best to avoid ultra-pasturization mlik products, and every lactose free milk is, aside from fairlife brand. Apparently they use a cold filtration technique so not only does this address the ultra-pasturatization issue it would also address your issue of there being no broken down remains of lactose from enzymes since the filtration just completely removes the lactose. So from what I gather this would be a good step in the right direction of good options to add to the list of how to go about making a probiotic product with the biogaia probiotic.
 

[TerryFirmer]

@Ovidus Most of your questions can be answered by carefully re-reading my previous posts.

 

Otherwise: Glucose water? Reuteri grows better if it has some protein to feed on. That's why the milk is heated to 90C - to de-nature (break up) the proteins because Reuteri is not great at proteolysis.

 

I take acacia fibre in an attempt to feed already resident gut bacteria other than Reuteri, the end products of which digestion will be food for Reuteri. I can't see any benefit to adding it to the fermenting medium. I take it at the same time as the Reuteri yoghurt.

 

20g glucose per L of milk.

 

 

[Ovidus]

Thank you TerryFirmer, 

One thing I am still unclear about after reading your prior posts is the following:

If we use lactose free milk and the product we have is one that has been pre-treated with lactase enzyme, the lactose in the milk will have already broken down into galactose and glucose, right? In that case, would the addition of glucose into the milk still be beneficial for the 6475 strain? 
I know previously you had said "extra glucose" so the above dynamics may have been considered already. However, I still wanted to ask.

 

Also: "I take acacia fibre in an attempt to feed already resident gut bacteria other than Reuteri, the end products of which digestion will be food for Reuteri."
So the other bacteria digest acacia fiber, and the end result of this digestion is food for the Reuteri? Do I get this right? Very, very interesting.....

 

Thanks again; I am learning a lot

 

Ovidus

I take acacia fibre in an attempt to feed already resident gut bacteria other than Reuteri, the end products of which digestion will be food for Reuteri. 

[TerryFirmer]

Thank you TerryFirmer, 

One thing I am still unclear about after reading your prior posts is the following:

If we use lactose free milk and the product we have is one that has been pre-treated with lactase enzyme, the lactose in the milk will have already broken down into galactose and glucose, right? In that case, would the addition of glucose into the milk still be beneficial for the 6475 strain? 
I know previously you had said "extra glucose" so the above dynamics may have been considered already. However, I still wanted to ask.

 

Also: "I take acacia fibre in an attempt to feed already resident gut bacteria other than Reuteri, the end products of which digestion will be food for Reuteri."
So the other bacteria digest acacia fiber, and the end result of this digestion is food for the Reuteri? Do I get this right? Very, very interesting.....

 

Thanks again; I am learning a lot

 

Ovidus

I take acacia fibre in an attempt to feed already resident gut bacteria other than Reuteri, the end products of which digestion will be food for Reuteri. 

 

Glucose favours 6475 over 17938, therefore it's beneficial to have more of it than is present in the milk after treatment with lactase. I have no idea what the upper limit of glucose utilisation would be, but in the papers I have read, 2% is a typical amount to add (ie 20g in 1L).

 

You are correct about my view on acacia. It contains rhamnose which is converted into 1,2 PD by gut bacteria. You could read chapter 4 of this thesis for the details:

https://era.library....d7-aa55f9834fde
 

[granmasutensil]

Glucose favours 6475 over 17938, therefore it's beneficial to have more of it than is present in the milk after treatment with lactase. I have no idea what the upper limit of glucose utilisation would be, but in the papers I have read, 2% is a typical amount to add (ie 20g in 1L).

 

You are correct about my view on acacia. It contains rhamnose which is converted into 1,2 PD by gut bacteria. You could read chapter 4 of this thesis for the details:

https://era.library....d7-aa55f9834fde
 

 

Terry came across this study while trying to find a way to isolate the two strains. It says reuteri specifically abundantly creates an antibacterial compound called reuterin when fed glycerin. Unfortunately I assume this won't help in isolating the 6475 and killing off the other strain since they are both reuteri. But maybe this is a really simple easy way to at the very least help prolong and maintain a dual culture and kill off any other strains that get in from batch to batch by simply adding say some small amount like 1/4 a teaspoon of glycerin each batch? Glycerin is super cheap and easy to get too. What do you think?

 

https://www.ncbi.nlm...les/PMC3365044/

[TerryFirmer]

Terry came across this study while trying to find a way to isolate the two strains. It says reuteri specifically abundantly creates an antibacterial compound called reuterin when fed glycerin. Unfortunately I assume this won't help in isolating the 6475 and killing off the other strain since they are both reuteri. But maybe this is a really simple easy way to at the very least help prolong and maintain a dual culture and kill off any other strains that get in from batch to batch by simply adding say some small amount like 1/4 a teaspoon of glycerin each batch? Glycerin is super cheap and easy to get too. What do you think?

 

https://www.ncbi.nlm...les/PMC3365044/

 

I didn't know that about glycerol, thanks. As you say, it won't help in splitting the two strains. (Actually, as a side note, 17938 produces much more reuterin than 6475.)

But you may be right that glycerol could help to retain batch purity. I'm pretty rigorous with keeping all vessels and utensils sterilized, so I'm not too worried about that. Also I use the same frozen whey from the first batch for all subsequent batches - I don't do serial inoculation. Once I run out of that, I start with tablets again.

[Ovidus]

Hi Terry Firmer, 

 

You obviously have a ton of information about the entire process. Do you work in a related field by the way?
Now, I just conducted my own little experiment, which I will share in the next post, but there is so much I am clueless about. For everyone's benefit, would you mind sharing your opinions on the below ? 
And since I am sure I neglected to ask many of the other pertinent questions, it would be tremendously appreciated if you can also point out any other matters yo consider crucial.

My Questions are: 

 

1- What kind of milk to use? Daily pasteurized milk that comes in a bottle (the kind which must be used in ~7 days or so) vs UHT... are both OK? Where I am, lactose free milk comes in the carton UHT only (does not need refrigeration before opening).
 

2- How to treat the milk before culturing? Bring to a boil? or just raise the temperature to 80 degrees Celsius? Shall we go beyond the point of sterilization to fully break up the protein bonds by keeping the milk at high temperatures for several minutes?

VERY IMPORTANT: CAN WE HEAT THE MILK IN THE MICROWAVE? 
 

3- With serial inoculation, the subsequent batches seems OK to the naked eye. Can you briefly explain why you prefer to freeze the original batch and use only that? I would have thought that if you have a method of creating an environment that is more friendly to the 6475 strain, subsequent batches would have a higher and higher concentration of just that strain, which would imply that serial inoculation would be preferred, no?

 

4- Can you explain how you make the original batch? Since it is not easy for me to get the tabs, I use just one crushed tab in  200 - 250 ml of milk and it takes a full 24 hours or even 30 hours to set. 

 

5- How much of the resulting yogurt do you consume and do you take it on an empty stomach? What is the timing of your acacia fiber intake? Do you take it some hours before the yogurt or does it not matter?

6- Can one consume the Reuteri yogurt forever or have you found (or have any theoretical reason to) that you may need to take a break occasionally?

 

7- Since regular folks won't have access to any testing, is there any basic rules we can use to assess if we are doing a good job preferring the 6475 over the other strain? (sweeter taste means...., harder vs softer texture means..... etc)

 

8- Since you are so well informed on this, by any chance are you making your own yogurt from any other strains too?

 

Many many thanks in advance. Your knowledge and contribution is tremendously appreciated. 

 

 

 

Edited by Ovidus, 14 September 2018 - 08:10 AM.

[Ovidus]

OK here is my little humble experiment: 

I made a starter batch first, using one tab in 200 ml of milk to produce the first cup of yogurt (200 ml of milk after it has been heat treated -so we are talking more "original" milk, since a good chunk gets evaporated; I keep it near boiling for 8 minutes or so)

 

I then used this batch to make 4 cups of yogurt

 

- Regular milk

- Regular milk + glucose (20 grams per liter)

- Lactose free milk

- Lactose free milk + glucose (20 grams per liter)

 

The results:

 

All batches were set when I checked them in ~ 16 hours time. With the original tab in pure milk (ie. the starter batch) in 16 hours you got just a little denser milk, and nothing at all that resembles yogurt. When I checked these at around that mark, all of them had the consistency of yogurt.

 

Another observation was that the 2 units with the glucose added were thicker. However, despite a good amount of glucose, their taste wasn't all that much sweeter, which is making me think that the Reuteri was consuming the glucose -at least to a large extent. 

[TerryFirmer]

@Ovidus Most of my knowledge came from reading this thread and all the various sources that other posters have linked to, plus searching Medline for relevant papers. I'm not a biologist and there are huge gaps in my understanding.

 

To try to answer your questions:

 

1. It's important to use lactose free milk, in whatever packaging it comes. I only have UHT skimmed or full fat available, and I have used both. Not surprisingly, the full cream milk gives a thicker consistency to the yoghurt.

 

2. I raise the milk to 90C for 20 minutes. Maybe ten minutes would be enough to denature the proteins, or maybe it needs 30 minutes - I simply don't know. I think you could heat it in the microwave, why not? I use a hob and stainless steel pan.

 

3. Good point about serial inoculation potentially increasing the percentage of 6475. I'll try it sometime. My current method is intended to minimise risk of contamination.

 

4. Please refer to my previous posts - I clearly described the process before. I used three tablets, figuring that at that rate one packet of 30 could last a year. The tablets are not easy to get where I live.

The very first batch, without a plain yoghurt co-culture, did take a very long time to set and there was a lot of separation of curds and whey..

 

5. I take the yoghurt three times per day, whenever I fancy it. 900mL lasts me five days. I take acacia with it once per day, usually with my breakfast shake. I've no idea whether consuming the two at the same time is preferable or not.

 

6. I see no reason to take a break. In fact, studies have shown that L Reuteri does not persist in the gut for more than about two weeks after we stop taking it.

 

7. No idea. We're fumbling in the dark. I asked a microbiologist friend and she said that telling the two strains apart would be enormously difficult. They would look exactly the same under a microscope, she thinks. We know that 6475 produces more histamine, 17938 more reuterin, but I've no idea how you would begin to measure those.

 

8. No, I'm solely focused on L Reuteri 6475.

 

As for your next post, it seems you're doing fine. Keep experimenting.

 

 

[Ovidus]

Tremendously appreciated Boss

You are the Man. 

 

Have you ever felt any histamine-related effects at all? 

[TerryFirmer]

Tremendously appreciated Boss

You are the Man. 

 

Have you ever felt any histamine-related effects at all? 

 

No, I've not felt anything unusual at all.

 

BTW I bought some casamino acids (not cheap at $100 for 500g) and I'm pretty disappointed with the results. I used 9g plus 9g of glucose in one 900mL batch and the yoghurt taste was strongly tainted by the yeasty/umami tang of the casaminos. Not revolting, but not pleasant, either.

 

So for my latest batch I used 4.5g casaminos and 13.5g glucose. The taste is much better.

 

The batches with casaminos are no thicker in consistency than those without, and the acidity is about the same (as far as my taste buds can tell) so goodness knows whether the L reuteri counts are higher (or lower).

[granmasutensil]

No, I've not felt anything unusual at all.

 

BTW I bought some casamino acids (not cheap at $100 for 500g) and I'm pretty disappointed with the results. I used 9g plus 9g of glucose in one 900mL batch and the yoghurt taste was strongly tainted by the yeasty/umami tang of the casaminos. Not revolting, but not pleasant, either.

 

So for my latest batch I used 4.5g casaminos and 13.5g glucose. The taste is much better.

 

The batches with casaminos are no thicker in consistency than those without, and the acidity is about the same (as far as my taste buds can tell) so goodness knows whether the L reuteri counts are higher (or lower).

 

Maybe the key isn't more macro nutrients since they already have a decent amount of glucose and fats and proteins from the milk but micro nutrients. Maybe adding a touch of something like a balanced multi mineral complex would be a good idea like coconut water which can be used for cell culture. But something without antibacterial qualities since I think coconut water has that.

 

I think this stuff might be a good idea. It is from prehistoric plant deposits which makes it different than just ionic minerals from evaporated sea water like the concentrace multimineral and the ratios of minerals I'm sure would be superior too. But it isn't so old that these plant deposits turned to coal. The mineral solution from these plant deposits has been used for cell culture and wouldn't have the antibacterial compounds coconut water has. It's also quite cheap and would last forever just for the yogurt making.

 

https://tjclark.com/...al-formula.html

 

If a person wanted to really go the extra mile this stuff is like the one previous mentioned but vastly improved and formulated specifically for the gut microbiome. It has other stuff like high end special metabolites that have been stabilized and stuff not listed on the label that makes it very special that a lot of money and research has been put into developing. I remember reading about it on a closed medical forum a few years ago and just remembered it and it's use in relation to this topic. Luckily it's sold in a small amount that would be perfect just to use for yogurt making. I'm probably gonna end up buying it next time I do a amazon order. I have no clue how much to use though...

 

https://www.amazon.c...store4life&th=1

Edited by granmasutensil, 16 September 2018 - 05:21 AM.

[Superball]

Hey guys, I am following this topic since some time. It is really really interesting what's going on in the last few pages and that more and more of you try to research and develop this idea.

 

For me, I also started making yoghurt from the Gastrus tabs and also came to the question how to seperate the two strains. I recently attended a bio hacking lab in my city and try to work out something with the guys. Right now we have a really promising idea but still have to work on growning the strains - in combination and seperately. 

 

What we found out - the only difference these two strains have is that 17938 grows on citrate, whereas 6475 does not. So the experiment I am setting up at the moment is the following:

 

1) I try to grow both strains on a petri dish (it seems harder than thought as the basic agar did not work and we are now trying a lactose agar) 

2) After that we copy (by stamping) the bacteria grown on this petri dish to a citrate agar petri dish and we should see that only 17938 grows on that

3) What we should have after that is 1 petri dish with both strains and 1 petri dish with just 17938 - so you should be able to spot 6475 by just looking at the dishes 

4) We would then pick up the 6475 bacteria and grow just those alone to get a starter for that

5) To check the validity of it we could then try to grow the extracted bacteria on citrate and see if anything grows - if nothing grows - we can be sure we isolated 6475

6) We could then even go on to multiply it and maybe freeze dry it to have a starter for the long run

 

What do you guys think about that? You think there is any mistake in the thought process? 


BTW got the info about the citrate from this paper: https://journals.plo...al.pone.0018783
https://s3-eu-west-1...242/preview.jpg
 

 

Also, as another idea and maybe a second experiment. While thinking about how to seperate the two - I had the idea that the tiny little spots (orange, red) on the tabs are not just decoration but could maybe be the growth medium of the two strains. Right now I am pretty sure that it is like that because if you look at the ingredients there are no things in it that could be that things (maybe apart from the armoa in it but nobody you put aroma in a tab in little spots that are not very well dissolvable + if you chew the tabs, these spots do not have flavour at all). 
So in theory, it could be possible that by simply seperating these little dots (and maybe knowing before if the orange or red ones are the 6475), washing them and incubating them seperately, one could extract the 6475 without a lab environment. 

Edited by Superball, 16 September 2018 - 07:30 AM.

[sdxl]

Superball, you should grow them on MRS agar. The only citrate agar I could find is not suitable for the growth of lactobacilli. You need to formulate your own agar if you want only citrate utilizing lactobacilli to grow on it. 

 

I have come up with my own plan to isolate ATCC PTA 6475. 

Inoculate MRS broth with a Gastrus tablet. Streak some of the cultured broth on MRS agar and incubate. Select a colony and streak onto fresh MRS agar and incubate. Repeat this step a few times. You need several plates for this to ensure at least one plate has a pure culture of  ATCC PTA 6475.

 

Prepare a broth with lactulose as the sole fermentable carbon source. Lactulose is not fermented by  ATCC PTA 6475 and quite well by DSM 17938. Inoculate the broth with a colony from a plate and inoculate. If the broth becomes turbid it is likely to be DSM 17938, if it stays clear it is likely to be ATCC PTA 6475. Alternatively you can make an agar of the broth.

 

 

BioGaia has answered the question on the dots on their website. 

 

What are the brownish dots that sometimes occure on the tablets?

The spots are not dangerous and the product can still be used. BioGaia tablets contains ascorbic acid, which is an antioxidant that contributes to the stability of the tablets. Over time the ascorbic acid oxidizes and the color becomes darker. This darkening is seen as small “spots” on the tablet.

 

[TerryFirmer]

@granmasutensil Do you have any links to peer-reviewed scientific papers showing that colloidal minerals increase the growth of Lactobacillae?

 

@Superball Props for thinking up an ingenious method to try to separate the two strains! But won't they be so homogeneously mixed together that it will be impossible to pick out separate colonies of 6475 with the naked eye?

 

@sdxl Sounds like a good plan. Hope it succeeds! Keep us posted.

[John250]

I just ordered the powder from Natures Way. 1 serving is 41.1mg
How much is everyone using?

[aribadabar]

I just ordered the powder from Natures Way. 1 serving is 41.1mg
How much is everyone using?

 

If your goal is to grow your own Reuteri ATCC PTA 6475, you bought the wrong product as it does not contain any of this strain.

Only Biogaia product Gastrus is known to offer it at the moment.

[Malf]

Okay I bought a pack of that Gastrus from iHerb and they are not like any other probiotic I ever took, first off they a Tablets, which is strange they are hard like a sweet tart candy and they say chew it.

 

I am very suspicious if this stuff works, I never heard of a probiotic put into a hard chewable tablet with orange flavoring

 

It seems also they only give you 100 million bacteria and usually you only get half of that so its 50 million if you lucky

 

They also charge an arm and a leg 26 bucks for a pack of 3 and such a small 100 million bacteria?

 

I took one today but ill finish the container and see if I feel any difference.

 

I started working out today as well dumbbells full body, Im going to work out 3 times a week with weights, and 2 times cardio, we will see.

[TerryFirmer]

@Malf and other recent posters:- Haven't your read the thread? There are dozens of posts here which would give you the info you require. Many of the posts here talk about how the Gastrus tablets are hopelessly weak (not to mention expensive) and thus we need to try to culture yoghurt from them. Several of us have successfully done so. Read up!

[Malf]

@Malf and other recent posters:- Haven't your read the thread? There are dozens of posts here which would give you the info you require. Many of the posts here talk about how the Gastrus tablets are hopelessly weak (not to mention expensive) and thus we need to try to culture yoghurt from them. Several of us have successfully done so. Read up!

 

Why do they make the tablets so weak? is it to make more money, I also read that the yogurt is dangerous to make at home if you are not a proper maker of this stuff.

 

Id rather just spend the money for more potent pill, is there anyway they could increase the probiotic count 100 is very low at least a billion, 100 million is really low, most common probiotics you can buy for cheap have 1 billion strain at least and many are 5 billion bacteria.

[Superball]

@Malf just give them a ring, I'm sure they can make an exception for you +46 (0)8 555 293 00

[John250]

If your goal is to grow your own Reuteri ATCC PTA 6475, you bought the wrong product as it does not contain any of this strain.
Only Biogaia product Gastrus is known to offer it at the moment.

Ahhh damn I ordered it because that was the brand recommended by selfhacked.

[Malf]

Ahhh damn I ordered it because that was the brand recommended by selfhacked.

 

I almost bought the one recommended by Self Hacked until someone posted about the Biogia Gastrus brand being the only one with the correct strain of L Reuteri

 

only downside is the Gastrus brand is so weak only 100million bacteria and also they put it in a chewable tablet, Ive wondered how the bacteria is alive if its mashed into a hard tablet, Ive only ever had bacteria in liquid capsules or capsules never hard tablet like that where you had to chew

 

they also add absorbic acid to it and citrus flavor.

 

it cost like 30 bucks as well

 

I hope another company comes out with this type of strain with more potency.

 

Ive only been on it two days and I will finish the entire box, ill see if I have any positive feelings or energy changes.

[John250]

I almost bought the one recommended by Self Hacked until someone posted about the Biogia Gastrus brand being the only one with the correct strain of L Reuteri

only downside is the Gastrus brand is so weak only 100million bacteria and also they put it in a chewable tablet, Ive wondered how the bacteria is alive if its mashed into a hard tablet, Ive only ever had bacteria in liquid capsules or capsules never hard tablet like that where you had to chew

they also add absorbic acid to it and citrus flavor.

it cost like 30 bucks as well

I hope another company comes out with this type of strain with more potency.

Ive only been on it two days and I will finish the entire box, ill see if I have any positive feelings or energy changes.

If 100million is weak what is optimal dose? Thanks

[Malf]

If 100million is weak what is optimal dose? Thanks

 

If you look on the market they sell different probiotic stuff that has 1 billion to 100 billion on them, and I read a lab study saying that half off the bacteria die off many times do to travel and storage conditions in hot or humid weather, so 100 million seems really low and how much of that is surviving?

 

What I don't understand is how the bacteria is able to survive in a tablet and it has brownish spots from the Vitamin C on the white tablets.

 

Some of the more powerful probiotics have billions of bacteria because they know most of it doesn't survive while on the shelf. and some will die off in your gut

 

Ill give it the full 30 before really giving a review, its only day two I took mine in the morning again right before eating breakfast.

 

This stuff is supposed to help upset stomach, increase regularity and help against constipation, that is the main description.

[Superball]

If 100million is weak what is optimal dose? Thanks

 

 

A good dose is likely the one used here. (at least it must have some therapeutic effects and should be kind of well researched if they rolled it in this study) 

 

 

 

orally receive 1010 colony‐forming units of L. reuteri 6475 daily

 

https://onlinelibrar...1111/joim.12805

 

 

So around 10 billion per day should be something to aim for. 

Edited by Superball, 08 October 2018 - 09:00 PM.

[John250]

A good dose is likely the one used here. (at least it must have some therapeutic effects and should be kind of well researched if they rolled it in this study)


https://onlinelibrar...1111/joim.12805


So around 10 billion per day should be something to aim for.

Yikes that would be 100,000 chewable tabs per day lol