448 replies to this topic

[hotbit]

Milk seems not a bad medium itself. The problem is that other strains are growing faster in milk than 6475.

I've done some calculations:
 

                                       strain A strain B

h                                     1.1            1.5

generation time h           18              18

Time – total h                   3               3

Time – lag ml                 200          200

Volume – total ml            5              5

Volume – initial billion/ml 0.02      0.02

Count – initial billion/ml 5.270      0.256

Count – in a jar  billion   1054        51    

 

Assuming growth is not inhibited (it is, but for simplicity of calculations I have assumed it is not) one tablet per 200 ml jar would yield CFU of 5 billion/ml  of one strain and 0.25 billion/ml of another one assuming the above ratio of generation times (growth rate / duplication time). If it would hold, one jar could have as much as ~50 billion of required bacteria, so not bad. However, there are many assumptions in the above calculations, so take it with a large pinch of salt The purpose is to show how easily one strain can out compete another one. The only way to tell how it works would be a lab test.

 

 

 I'm thinking of buying a yogurt maker. Can the temp be controlled or is it set at a certain temp?

 

Some  can, some cannot. I bought like this one but branded differently. Temp. is not very uniform, i.e. jar in the centre is warmer 2-4 degrees than those outside. I set it up at 35C to have 37-38 in jars. (Oh, it's months since I have used it...)

Edited by hotbit, 25 November 2017 - 12:57 PM.

[Liminal_God]

I came across one of the studies on the 6475 strain a couple weeks ago and immediately became interested in supplementing it.  I bought the Gastrus tabs and just this past weekend made my first batch of fermented milk with it.  It was definitely a success and I will try to post some pictures and steps I took of the process within the next few days.  I've done 3 days of 1 tab/day and then 4 days of 1/2 C milk/day. So far I haven't noticed any apparent effect other than maybe improved bowel movements.

 

 

[ekaitz]

Been thinking; as for the concern about milk strains outcompeting the 6475.. what if, instead one tab you put 1 or 2 full packets into the milk? that way chances are higher the 6475 will "rule" the milk strains, right?

Edited by ekaitz, 13 February 2018 - 10:33 PM.

[holdorfold]

Dr. William Davis has written an article in April on undoctored.com about making yogurt with the BioGaia Gastrus.

 

Quote:

 

"Problem: There are only 100 million CFUs (live organisms) per tablet. I have not observed any substantial health benefits by ingesting the tablets.

 

So I have been amplifying bacterial counts by making yogurt. The counts are further increased by performing fermentation in the presence of prebiotic fibers. Just as ingesting prebiotic fibers increases bacterial counts in your intestines, so it goes in making yogurt, as well.

 

The yogurt is thick, delicious, and contains a marked increase in bacterial counts, though I have not yet performed a formal count. Given the extraordinary thickness of the end-product, it is likely that trillions of CFUs are present...

 

People who consume 1/2 cup per day of this preparation (mix with blueberries, strawberries, etc.) are reporting the effects listed..."

Full article here: https://blog.undocto...reuteri-yogurt/

 

*Edit* He's also posted this article informally examining subsequent batches of the yogurt under a microscope: https://blog.undocto...reuteri-yogurt/

Edited by holdorfold, 04 June 2018 - 05:18 PM.

[sdxl]

Being a cardiologist does not make you automatically knowledgeable in microbiology. I have read all of his blog enteries on this "yogurt" and I'm not impressed. MD or not he should substantiate his claims. Either by citing relevant studies, or conducting his own tests.

 

He is making assumptions on how inulin increases the growth of the bacteria. The strains in Gastrus do not ferment inulin, as shown in the study posted here earlier. Even when inulin is fully hydrolyzed by other micro-organisms in vivo, they can barely ferment the fructose. I expect at least that he did a plate count of one batch with and one without inulin with to support his claim. 

[holdorfold]

I've since made two different batches of the stuff, the second batch turned out much better than the first. I use goats milk instead of cow's milk because it's less estrogenic and I used inulin in one batch and left it out in the other.

 

First batch I used 1L of goats milk, 1 tablespoon of inulin and 9 crushed Gastrus tablets and left in a yogurt maker for 24 hours. The result was very runny, the consistency of the goats milk hadn't thickened at all, but apparently goats milk doesn't really thicken much for yogurt making anyway. It didn't taste very sour, there was still sweetness to it, so I assumed it didn't ferment a huge amount or perhaps the inulin hadn't fermented in particular. Upon ingesting about half a cup, I felt a noticeable sense of relaxation so I laid down for a bit and noticed that my bed felt more comfortable, my sense of tactile appreciation of it had increased. I assume this is to do with increased oxytocin levels produced by one of the strains.

Second batch, after reading the above reply, I left out the inulin. I used 1L of goats millk and this time added 300g of goats cream to thicken it a bit. I added about half a cup of the previous batch as starter. Left in the yogurt maker for 24 hours. This time it had separated into liquid on the bottom and solid on top, which should indicate a stronger fermentation. I stirred it together and it was much, much thicker than the previous batch. With the previous batch, the consistency of hadn't noticeably changed before and after fermentation, this time it  had changed a lot, so it wasn't just the extra cream. The taste was a lot more sour as well, and this time, I got a strong histamine reaction to it after ingesting. I basically start sneezing after having half a cup of this stuff, this has happened every time I try it. Apparently the strain increases histamine production, so it seems to be a positive indication. I still get the relaxed feeling after drinking as well. Note: I didn't get the sneezing from the gastrus tablets alone or the first batch, so I assume it has something to do with increased bacterial count in the second batch.

Edited by holdorfold, 17 June 2018 - 11:05 AM.

[sdxl]

I made a broth and inoculated with a single Gastrus tablet for 24 hours.

 

 Composition of 1 liter (in grams): 

  

 Glucose                            20

 PeptoPro                           15

 Yeast extract                    11.5

 Sodium acetate                      5

 Potassium citrate monohydrate     2.6

 Diammonium phosphate              1.6

 Sunflower lecithin                  1

 Magnesium sulfate heptahydrate    0.2

 Manganese sulfate tetrahydrate   0.05

 

The medium became turbid. As expected gas was produced. Didn't screw the cap on too tightly to let the carbon dioxide escape. An airlock might be useful. It isn't the most palatable thing you can drink, but I'm sure there are much worst tasting things out there.

  

The yeast extract I used contains 48.6% protein and 18.5% carbohydrates and is low in salt. Yeast extracts can vary quite a lot. The only powdered yeast extract I could find has way too much salt, so this will have to do. It is a real pain to measure this dark brown, thick, sticky, smelly paste.

 

Sunflower lecithin replaces polysorbate 80. See US patent 9,468,648 for further details. In theory, as an oleic acid source it may also enhance gastric survival of lactobacilli.

 

I did try to adhere to the recipe for MRS as much as possible with the limited resources I have. I omitted the beef extract, so I increased the PeptoPro and yeast extract. In no way this is an optimized medium, because that would require a decent lab setup and a lot of testing. I likely will be adapting the medium to make it more like the one in the patents, which is nutrient richer.

Edited by sdxl, 19 June 2018 - 09:27 PM.

[sub7]

Question to all: 

Here in Europe, I was able to find a few products that claim to contain Lactobacillus Reuteri, but -at least on the package itself- there seems to be no specific information about the exact strain (maybe in the package insert there is, who knows). 

Are there several sub-strains of Reuteri ? Or if a serious company is labeling a culture as Lactobacillus Reuteri, can we safely assume that it is ATCC PTA 6475?

 

 

[holdorfold]

Are there several sub-strains of Reuteri ? 

 

 

Yes, there are many different strains of Reuteri.

 

 

Or if a serious company is labeling a culture as Lactobacillus Reuteri, can we safely assume that it is ATCC PTA 6475?

No, it's a very rare strain to find in a probiotic, infact, I believe BioGaia Gastrus is the only product on the market which contains this strain since I think the company has patented ATCC PTA 6475.

[sub7]

thanks a lot for the great response...

[Moondancer]

I took Biogaia's product the past months and haven't noticed any effects either. I agree the bacteria count of the product seems to be very low. On the other hand, when you culture it in a yogurt as someone suggested the second bacteria strain may as well start to rule over the strain you want to culture. I do sometimes culture a single bacteria strain probiotic in yogurt, but how can you be sure you are not ending up with a product mostly containing the second strain and with hardly any LB ATCC PTA 6475 (if any at all), once you start to culture a probiotic with 2 strains in yogurt?

 

 

[jolly]

Any downside to keeping the tablets in the fridge? 

[holdorfold]

how can you be sure you are not ending up with a product mostly containing the second strain and with hardly any LB ATCC PTA 6475 (if any at all), once you start to culture a probiotic with 2 strains in yogurt?

 

According to this blog post: Histamine Reaction to BioGaia Gastrus, several people report increased histamine symptoms upon consumption of the Gastrus tablets. Apparently, the mode of action of specifically ATCC PTA 6475  is to generate histamine which activates H2 receptors in the gut. 

So we have a symptom here which should be unique to that particular strain and not the other one. I don't get any histamine response from just the Gastrus tablets, but I do get a strong histamine response from the cultured yogurt which should indicate a greater presence of the 6475 in particular.

 

In addition to that, there are many anecdotal reports in the comments section of Dr. William Davis' blog posts about making the yogurt where people are saying they are achieving the desired effects which relate to the 6475. But apart from these anecdotal reports and my case, I guess one would need to do a proper lab test on the cultures on be completely certain.

[sdxl]

The only way to ensure that only ATCC PTA 6475 grows in your medium is to start with a pure culture. You've got these options:

 

 Buy it from the ATCC. 

 

 Isolate it from Gastrus. 

 

 Wait until BioGaia releases a single strain product with it.

 

 

Buying it from the ATCC requires a certification and is expensive. The first two options require microbiology skills and a microbiology lab. I'm assuming most of you don't have either, so the last option remains.

[sdxl]

Any downside to keeping the tablets in the fridge? 

 

No, they should maintain their potency longer refrigerated. 

[sub7]

OK Gentlemen, 

 

Reporting on my experiment with a different Reuteri strain. I just got my hands on tabs of Lactobacillus reuteri Protektis (L. reuteri ATCC 55730) and tried making a yogurt with it. I did not even have a thermometer at home and -as a first go- I conducted a very simple experiment involving the following steps: 

 

1- Bring raw milk to a very mild boil and as soon as boiling occurs turn down the heat and keep at at around that temperature for another ~ 5 minutes.

 

2- Let it cool down so that when you insert your finger into a sample of the milk (in a separate cup, so this is done without contaminating the actual lot) is tolerable and won't burn you.
 

3- Crush 3 tablets of the above strain of Reuteri (each tablet is claimed to have 100 million bacteria) and add the resulting powder toa teaspoon of sugar. Mix it all into around 300 ml of milk prepared in steps 1&2.

 

The tablets were made by Biogaia btw and were sent by a friend who got them for me in his country. No refrigeration needed on the tabs, while the liquid form was best refrigerated. 

 

 

After around waiting for 6-7 hours (wrapped up in a blanket to keep warm), the mixture prepared as above had attained no nigher consistency than plain milk. So I drank some and let the rest sit on the counter -this time not even wrapped up or anything, just let it sit at room temperature. Amazingly, however, after another 10 or so hours, it turned into a pudding and tasted as such. The sugar therein overpowered the taste of any sourness (if such was present at all) and the whole thing had a pretty tolerable taste. 
I was surprised that such a causal approach resulted in (slightly watery) yogurt. Not only was the approach very haphazd as you can see, but also the strain itself was not advertised as a yogurt starter. So very encouraging results overall, as it suggests that at least multiplying the bacteria count is fairly easy -with this particular strain anyways. 

 

Now, I understand that the strain ATCC PTA 6475 is supposed to be the best among the various types of Reuteri. But any idea at all (or rather any speculation at all) how much of the similar benefits we can derive from the above mentioned variety of Reuteri?

 

[holdorfold]

Now, I understand that the strain ATCC PTA 6475 is supposed to be the best among the various types of Reuteri. But any idea at all (or rather any speculation at all) how much of the similar benefits we can derive from the above mentioned variety of Reuteri?

 

The effects we're after from the 6475 were discovered through studies on rats using that particular strain and not others. We can only guess if other strains might have similar effects, there's no data on that at the moment.

The closest anecdotal report I've heard from someone having 6475-esque effects from other strains was under the comments section of a blog discussing L. Reuteri for Hair Loss: https://perfecthairh...ir-loss-review/

The comment reads: "I found Nature’s Way Primadophilus on Amazon and started in in March or April. I did not measure the boys [testicles] until May, I use a caliper. Since May, one guy has grown 9% in length, the brother, 23%!"... and he remarks: "While there may be differences in this effect, or that effect, I suggest that most strains will give you the benefits you want."

 

The product he is talking about is here: https://www.amazon.c...c/dp/B000FL9EAG

 

It contains 5 Billion CFU which is a lot more than Gastrus' mere 200 million CFU, but of course it doesn't have that golden studied strain.

So perhaps if you're making yogurt from a different strain using a BioGaia product, you might as well just use the Nature's Way product as is since it contains 5 billion CFU, so you don't have to increase anything through culturing. Either that or try with the BioGaia Gastrus which contains the strain which studies have shown have the effects we're after.

I'm going to buy the Nature's Way product I think to test on myself to see what effects it has, perhaps at higher doses than recommended as well.

[Benko]

I can't paste links into this form from this computer so you'll have to google it, but there was a recent study (also animals) abut R6475 being beneficial for bone density.

 

FWIW I'm an atypical case as my gut flora was messed up by daily penicillin from age 3-13, but I did notice a significant difference e.g. regular bowel movements taking R 6475.  The chewable tabs bio..... brand.

[sdxl]

I believe you're talking about this study.

[TerryFirmer]

I have started making yoghurt with the Gastrus strains. I'm using lactose-free cows' milk because lactose favours growth of the wrong strain. The lactose is split into glucose (which favours 6475) and galactose (which favours the other, but not as strongly as lactose, and there's 50% less of it). I heated the milk to just below boiling and kept it there for about ten minutes, then let it cool to about 40 degrees C. To 900mL of the full cream milk I added 20g glucose and 5g arabinogalactan fibre, plus three Gastrus tablets crushed up.

 

A cheap Chinese yoghurt maker holds the milk at about 41 degrees C. The first batch took 30 hours to fully thicken, and the gas inside made it almost overflow out of the steel container inside the yoghurt maker. There was strong separation between the yoghurt and the whey, the former floating on top, looking more like a very yeasty bread dough in texture. I skimmed it off for cooling and consumption. The whey was put into my ice cube trays for use as future starter. I also kept a little in the fridge for the second batch.

 

I used about 50mL of the whey for that, together with 20g glucose but no arabinogalactan this time. I tried skimmed lactose-free milk, prepared in the same way as before. The yoghurt thickened in just 8 hours. It has a much more even, smooth consistency with far less separation than batch #1. I was able to stir most of the whey into it without making it too runny.

 

Batch #1 tastes rather sweet, so maybe I put too much glucose in it. Batch #2 is less sweet. It's far too early to tell if there will be any health benefits, but I'm pleased to have got the process under way.

[TerryFirmer]

I just found this patent: https://patents.goog.../US7344867B2/en

 

If I'm reading it correctly, the experiments described indicate that 55730 (the parent of 17938) is pro-inflammatory, leading to increased TNFα production in vitro:

 

 

Incubation of THP-1 cells with 24-hour L-CM from L. reuteri ATCC PTA 4659, L. reuteri ATCC PTA 6475 strains in the absence of LPS did not lead to the generation of TNFα in the incubation medium (FIG. 1). Surprisingly, L-CM from L. reuteri ATCC 55730 and L. reuteri strain CF48-3A stimulated the production of TNFα by the THP-1 cells to levels similar to those seen with LPS alone. Thus, the L. reuteri strains differ in their ability to stimulate pro-inflammatory TNFα production by resting THP-1 monocytes.

Addition of LPS to the THP-1 cells in the absence of L-CM led to the generation of 138 pg/ml TNFα during the 3.5 hour incubation period. This is the expected inflammatory response of the THP-1 cells to the toxin. Addition of the growth medium (MRS), which acts as a control for the L-CM additions, led to the generation of 132 pg/ml TNFα and thus MRS did not interfere with the response to LPS. The addition of 24-hour L-CM from L. reuteri ATCC PTA 4659 or L. reuteri ATCC PTA 6475 dramatically reduced the levels of LPS stimulated TNFα to only 14 and 10 pg/ml, respectively. This represents an inhibition of LPS-stimulated TNFα production of 89 and 92%, respectively. On the contrary, in the presence of 24-hour L-CM from L. reuteri ATCC 55730 and L. reuteri strain CF48-3A, LPS was still able to induce a significant rise in TNFα compared to the levels in the absence of LPS. LPS-stimulated TNFα production increased by 50% and 38% despite the presence of L-CM from L. reuteri ATCC 55730 and L. reuteri strain CF48-3A, respectively.

Similar experiments performed with 9-hour L-CM from L. reuteri ATCC PTA 4659 or L. reuteri ATCC PTA 6475 demonstrated that the inhibitory effect on LPS-stimulated TNFα production was considerably less (52% and 16%, respectively; FIG. 2) but still there. Thus, longer incubations of the L. reuteri strains, with harvesting of the L-CM in late log/stationary phase of growth, leads to improved efficacy in inhibiting TNFα production.

.....................................................

Compared to medium controls, the production of TNFα in response to LPS in differentiated macrophages derived from primary monocytes from CD-act patients, was significantly inhibited by 50% by L-CM from L. reuteri ATCC PTA 4659 and by 60% by L-CM from L. reuteri ATCC PTA 6475 (FIG. 4). Confirming the data from the THP-1 cells, L-CM derived from L. reuteri ATCC 55730 and L. reuteri strain CF48-3A were unable to inhibit the production of TNFα and on the contrary, increased the production of TNFα by 22% and 30%, respectively compared to the relevant controls

 

Assuming that the genes that were knocked out in creating 17938 from 55730 (genes to do with potential antibiotic resistance) have not altered the above characteristics, I'm really puzzled why BioGaia combined an anti-inflammatory (6475) and a pro-inflammatory (17938) in the same product.

 

Anyway, it further emphasises the importance of adjusting the culture medium to strongly favour 6475 by any means possible.

 

Edited by TerryFirmer, 06 August 2018 - 10:53 AM.

[Mike C]

I’m 65 and have been reading stuff like this all my life. Remember Adele Davis? I read her books ha ha! The fountain of youth search, never ends!

[Izan]

Why haven't they released a super potent stand alone atcc pta 6475 product yet?

 

 

[John250]

Anyone have a link for the most current best brand selling it in capsules?

Edited by John250, 24 August 2018 - 07:48 PM.

[sdxl]

Why haven't they released a super potent stand alone atcc pta 6475 product yet?

 

You should ask BioGaia. BioGaia's competitor Probi is going to launch a probiotic for bone health this year. Since the ATCC PTA 6475 probiotic will be competing in the same new area BioGaia can't stay behind long.

[Moondancer]

I just found this patent: https://patents.goog.../US7344867B2/en

 

If I'm reading it correctly, the experiments described indicate that 55730 (the parent of 17938) is pro-inflammatory, leading to increased TNFα production in vitro:

 

 

Assuming that the genes that were knocked out in creating 17938 from 55730 (genes to do with potential antibiotic resistance) have not altered the above characteristics, I'm really puzzled why BioGaia combined an anti-inflammatory (6475) and a pro-inflammatory (17938) in the same product.

 

Anyway, it further emphasises the importance of adjusting the culture medium to strongly favour 6475 by any means possible.

 

Interesting. I agree with those members that said BioGaia Gastrus is a too low dose probiotic.

 

Anecdotally I have not noticed anything whilst taking it the past year - I have wondered occasionally if it could be worsening my autoimmune symptoms, but I have nothing really to base that conclusion on. I don't feel better or worse when I don't take it. But your suggestion that the second strain would have pro-inflammatory properties, does worry me. I may stop taking it, since I haven't noticed any benefits while taking it either way.

[granmasutensil]

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?

[TerryFirmer]

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?

 

Without bacterial counts we're fumbling in the dark. All we really know is that lactose is likely to favour the growth of 17938, so lactose-free milk is better than ordinary milk, and extra glucose should favour 6475. I've no idea about B vitamins.

 

I have some casamino acids on order, which certainly greatly help 17938 to multiply and I'm hoping the effect extends to 6475 also.
 

My batches of yoghurt are consistent in texture and taste, but it's too early to see any health effects. I'm taking 140mL a day - should be considerably more powerful than three Gastrus tablets. But again, I don't know how much of each strain there is.

 

It's also important to feed the other gut bacteria whose end products feed the L Reuteri in the large bowel. To that end I'm taking acacia fibre which has a decent amount of rhamnose which in turn should result in production of 1,2 PD by B. Breve and other common gut bugs.

Edited by TerryFirmer, 28 August 2018 - 01:11 PM.

[ekaitz]

Do you think is best to cultivate gastrus strains by simple yogurth making, or is preferable to use kefir nodules?

Edited by ekaitz, 05 September 2018 - 12:13 PM.

[TerryFirmer]

Do you think is best to cultivate gastrus strains by simple yogurth making, or is preferable to use kefir nodules?

 

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