448 replies to this topic

[Heisok]

Great update hotbit. Thanks.

 

What I assumed that I lose during the straining through coffee filters is the liquid which is clear with a greenish/yellow tinge. Figured mostly the Galactose, and whey with some of the bacteria. Since the yogurt works better than the first batch in the subsequent batches using some separately saved yogurt, I figure that at least a good concentration of the bacteria is present. Then again, I do not strain off all the liquid, so it might simply be enough whey left to have some bacteria. I went as far as 4 generations, but for 5th and 6th, I added a few Gastrus to supplement the saved Yogurt as a starter.

 

I am also encouraged that my method should retain a large amount of bacteria from the claims of White Mountain Foods. They also use a 24 hr ferment, which for the Greek version is strained. They claim 250 Billion CFU per 8 ounces.

 

http://whitemountain...eek-yogurt.html

 

I miss my Yogurt. Time flies, it has been 6 months since I last made it. What I can say is that I am probably about 8 pounds heavier eating a similar very low carb diet without the Yogurt. I think my replacement calories are coming mostly from fats.  As slightly off topic questions, is there an opinion about if Casein is a protein which can be harmful? Is it true that whole milk can be Estrogenic?

 

One additional thought. I thought that maybe since L.Reuteri strains can grow on Galactose, that it is maybe part of the reason that adding Yogurt bacteria which create Galctose by consuming Lactose, to the incubation helped multiply them over the culturing broth or milk.

Edited by Heisok, 27 February 2017 - 11:42 PM.

[hotbit]

As slightly off topic questions, is there an opinion about if Casein is a protein which can be harmful? Is it true that whole milk can be Estrogenic?

 

Well, I'm not an expert in the field. Leaving casein aside, there are cow hormones, as I know mostly present in fat, including estrogen. Still, it may not necessary be harmful. Probably nobody knows...

https://www.elsevier...oductive-health

 

On the side note, these two articles might be of your interest?

https://www.scienced...70223124259.htm

https://thequantifie...mimicking-diet/
 

 

PS

''They claim 250 Billion CFU per 8 ounces.'' ~ 0.9 billion per ml is quite excellent.

Edited by hotbit, 28 February 2017 - 12:09 AM.

[fluidity]

I believe I recall one of you guys using natren probiotics earlier in the thread. Can anyone else comment on this? I'm curious if it's worth trying out as well

[Black Fox]

Hi guys? Who are you guys buying the strain from? Has any of you tried with coconut milk? ( cuz I'm intolerance to dairy

I've been successful in making my own spermidine yoghurt with coconut milk.

[orion602]

what about using goat or sheep milk for making a yoghurt, would it make any difference?

 

 

I've been successful in making my own spermidine yoghurt with coconut milk.

 

making spermidine yoghurt? how?
 

Edited by orion602, 20 May 2017 - 09:44 PM.

[Skyisred]

Is there a way to culture it without dairy ? I can't have any, including goats and sheep milk. Also cows milk does have a lot of hormones which our bodies do not need. Consuming extra growth hormones in adulthood have been linked to cancer.

Edited by Skyisred, 14 September 2017 - 09:08 AM.

[sdxl]

Is there a way to culture it without dairy ? I can't have any, including goats and sheep milk. Also cows milk does have a lot of hormones which our bodies do not need. Consuming extra growth hormones in adulthood have been linked to cancer.

 

Yes, it certainly possible to grow the bacteria on a vegan medium as long as their needs are met.

[Skyisred]

Could you elaborate please? Any specific mediums to grow it?

Edited by Skyisred, 14 September 2017 - 05:34 PM.

[sdxl]

Could you elaborate please? Any specific mediums to grow it?

 

You have to know what the bacteria need and provide it to them. The standard medium for growing lactobacilli is MRS (de Man, Rogosa, Sharpe) broth or agar. I suggest to study all the ingredients in depth to understand what the bacteria need to grow. The peptone in MRS can be derived from milk protein. This can be replaced by hydrolyzed protein from soy, pea, etc. Any modifications should be tested thoroughly.

[hotbit]

Could you elaborate please? Any specific mediums to grow it?

 

You can try in coconut (can be diluted) milk with some Vit B added and a spoon of glucose or maltose. I had some mixed results trying cultivation in normal milk. Difficult to say what is happening if unable to check / count bacteria in a sample. Maybe tablets were not good, maybe I have just misinterpreted results. Just test medium and ideas you have. Find what works, share your results.

[sdxl]

 

Could you elaborate please? Any specific mediums to grow it?

 

You can try in coconut (can be diluted) milk with some Vit B added and a spoon of glucose or maltose. I had some mixed results trying cultivation in normal milk. Difficult to say what is happening if unable to check / count bacteria in a sample. Maybe tablets were not good, maybe I have just misinterpreted results. Just test medium and ideas you have. Find what works, share your results.

 

 

Anything other than glucose as a fermentable carbohydrate is not recommended if you are using the Gastrus tablets to grow ATCC PTA 6475. In a previous post of me, you can see the strain of which DSM 17938 was derived from grows more than twice as well in a medium containing maltose than ATCC PTA 6475. Same goes for a medium containing lactose, so culturing it in milk doesn't make sense.

 

Be aware that Lactobacillus reuteri produces carbon dioxide and could make your container explode if the gas can't escape.

[TerryFirmer]

Great thread. How are the cultures coming along, guys - any new tips?

 

I see some comments about culture medium in this paper: https://www.ncbi.nlm...les/PMC4825993/

 

"L. reuteri ATCC PTA 6475 was initially cultured on deMan, Rogosa, Sharpe media (MRS, Difco)—agar plates and kept under anaerobic conditions at 37°C for a maximum of 1 week. For gavaging of all mice, one colony of L. reuteri was picked and anaerobically cultured in 10ml of MRS broth for 16-18h at 37°C. MRS broth was additionally used as a negative control. For drinking water, L. reuteri was anaerobically cultured in 10 ml of MRS broth for 16-18h at 37°C. The following day, the overnight culture was sub-cultured into fresh MRS and grown until log phase (OD₆₀₀ = 0.4). L. reuteri was spun down, re-suspended in sterile PBS, cfu/ml calculated and stored at -80°C until use. L. reuteri was re-suspended in drinking water at a final concentration of 3.3x10⁸ cfu/ml."

 

Seems like 37 C is right on the money. Anaerobic culturing sounds tricky, though.

[joelcairo]

I recently made some yogurt using Swanson's L. Reuteri product, which actually contains:

Lactobacillus Reuteri - 5 billion
Lactobacillus Rhamnosus - 1 billion
Lactobacillus Acidophilus - 1 billion
 

I cultured it in a loosely covered bowl and used the oven light method to keep it at around 37 degrees C. I took it out after about 16 hours, which was probably a few hours too long. It looked runny in the oven, but after going into the fridge it firmed up quite a bit.

 

A small amount of liquid separated out, with a greenish tinge to it similar to pictures posted earlier in the thread, but much less liquid and I just stirred it back in.

 

I can confirm that it improved the end products of my digestion a little bit but I didn't become aware of any other effects or benefits.

 

I'm on a keto diet right now, to lose weight, so I am going to continue playing with these yogurt strains after I go back to a more normal diet. I think the high fat and low carb nature of the diet is going to skew my intestinal bacteria more than any yogurt will.

Edited by joelcairo, 19 October 2017 - 08:06 PM.

[ta5]

I recently made some yogurt using Swanson's L. Reuteri product, which actually contains:

Lactobacillus Reuteri - 5 billion
Lactobacillus Rhamnosus - 1 billion
Lactobacillus Acidophilus - 1 billion

 

A little over a year ago I asked Swanson which strain of Reuteri was in their product, and this was their response:

 

We appreciate your patience while we further investigate your inquiry. After speaking with a knowledgeable Product Specialist, they informed us that we are not provided with the specific strain of the Swanson Probiotic L. Reuteri Plus item #SWA025. Also, please know that the specific strain can vary with each batch produced. We hope this information is helpful. Please contact us if you have further questions.

Our quality, value and customer service have made Swanson Health Products a trusted source of nutritional supplements since 1969.  To help us build this privileged relationship with you, we welcome your comments and suggestions, as it is our pleasure to serve you at any time.

Have a Happy & Healthy Day!

Kari D.

Swanson Health Products

Customer Account Specialist

customercare@swansonvitamins.com

www.swansonvitamins.com

 

I never bought it again.

[TerryFirmer]

@ta5 Yeah, if it's not the exact correct strain (6475 in this case), it's probably a waste of time. I'm pretty amazed that Swansons don't know what they use to make their own product.

[holdorfold]

I'm going to purchase BioGaia Gastrus and want to make yogurt out of it to increase the supply. This is something I've never done before.

Since I live in shared accommodation I can't use the oven for too long so I'm planning on buying a cheap yogurt maker like this one which yields one liter.

Is it just a matter of putting milk in it and a crushed Gastrus capsule, or do I need to add more ingredients to make it work?

Thanks

Edited by holdorfold, 28 October 2017 - 10:17 PM.

[sdxl]

You can't just dump a probiotic into milk and hope for the best. There are journals devoted to fermenting milk. Milk is not an ideal medium for these bacteria. It requires the inclusion of other bacteria to make milk a more suitable medium by hydrolyzing proteins, etc. 

 

Milk seems a more ideal medium for DSM 17938 than for ATCC PTA 6475, because it significantly grows better in a lactose containg medium. Or at least the strain from which it was derived, which isn't that different. Don't ferment Gastrus tablets in milk if you want ATCC PTA 6475 to thrive over DSM 17938. If you happen to have a pure culture of ATCC PTA 6475, you could experiment with including other cultures. This requires careful selection.

    

Crushing the Gastrus tablets is unnecessary and may contaminate the culture. The tablets will dissolve.

 

This topic is way too obsessed with making yogurt, not with optimizing the growth of the strain.

 

 

Anyway, my DIY broth would include:

 

Glucose - fermentable carbohydrate. 

PeptoPro (DSM) - hydrolyzed casein. Source of peptides, low lactose.   

Yeast extract - all sorts of stuff for the growth of lactobacilli. 

Lecithin - replacement for polysorbate 80.

Epsom Salt - magnesium source

Other food grade chemicals to match original MRS.

Other ingredients that show to increase the growth of lactobacilli universally

  

[holdorfold]

Thanks for the info.

 

 

 

You can't just dump a probiotic into milk and hope for the best. There are journals devoted to fermenting milk. Milk is not an ideal medium for these bacteria. It requires the inclusion of other bacteria to make milk a more suitable medium by hydrolyzing proteins, etc. 

 

Milk seems a more ideal medium for DSM 17938 than for ATCC PTA 6475, because it significantly grows better in a lactose containg medium. Or at least the strain from which it was derived, which isn't that different. Don't ferment Gastrus tablets in milk if you want ATCC PTA 6475 to thrive over DSM 17938. If you happen to have a pure culture of ATCC PTA 6475, you could experiment with including other cultures. This requires careful selection.

 

 

L Reuteri 6475 is in breastmilk so you'd think milk would be a descent medium for it.

 

Do you have any sources for 17938 out-competing 6475 in lactose containing mediums?
 

Cheers

Edited by holdorfold, 01 November 2017 - 06:02 AM.

[holdorfold]

On a side note, I've just found another product which claims to have the 6475 strain in it.

It's called Floracil50 by Truthnutra and contains 2 Billion CFU's of L-Reuteri which according to this video: https://youtu.be/n8Rh7PnJbTA?t=6m54s is the 6475 strain.

 

 

 

 

 

Edited by holdorfold, 01 November 2017 - 06:31 AM.

[sdxl]

 

L Reuteri 6475 is in breastmilk so you'd think milk would be a descent medium for it.

 

Do you have any sources for 17938 out-competing 6475 in lactose containing mediums?

 

 

The strain can ferment lactose. Proteolytic activity might not be sufficient for prolific growth in milk, as is the case with many probiotic lactobacilli. Should be tested to be certain.

 

DSM 17938 is derived from  ATCC 55730. I have no indication that the fermentation profile is significantly different between the two strains. 

The study  I was referring to compares ATCC 55730 and ATCC PTA 6475.  (see figure S7)

[sdxl]

On a side note, I've just found another product which claims to have the 6475 strain in it.

It's called Floracil50 by Truthnutra and contains 2 Billion CFU's of L-Reuteri which according to this video: https://youtu.be/n8Rh7PnJbTA?t=6m54s is the 6475 strain.

 

 

 

 

If it does contain the strain this would be patent infringement. You would have to take their word they expressed in a poorly made video. Why are none of the strains used in this supplement listed on the label? I would never consider buying this supplement.

[recon]

Is there any supplement containing just ATCC 6475?
Even Gastrus contains another Reuteri, I believe. Is there not one on he market just having that one particular strain?

[mkmicro]

I've been following this thread and decided to try my take in testing the Gastrus for bacteria. I crushed the tablet into powder and did a test in 4 ounces of whole milk @ 37 degrees Celsius in a glass jar with a cloth covering the top (for the carbon dioxide to be released) for 20 hours. My main point in testing was to make sure there was live bacteria. I had a decent result. Top of liquid was soft, almost like yogurt and middle had inoculate (cloudy mixture). I did a quick taste test and it tasted like unflavored Kefir. I am thinking of doing another test and boil the milk first (as I read earlier). Should I use raw milk or just the same store bought whole milk I used the first time? Any suggestions? Also, I put the jar in the fridge and the next day it is quite sour - much more than the day before. Btw, very interesting thread!

[holdorfold]

 

 

If it does contain the strain this would be patent infringement. You would have to take their word they expressed in a poorly made video. Why are none of the strains used in this supplement listed on the label? I would never consider buying this supplement.

 

 

Yeah I'm also dubious about it, I've sent them an email requesting more information and am awaiting a response. Will update when I hear back from them.

Is there any supplement containing just ATCC 6475?
Even Gastrus contains another Reuteri, I believe. Is there not one on he market just having that one particular strain?

 

Not that anyone knows of. Infact, the only one I'm aware of that definitely contains 6475 at all is the Gastrus product.

[holdorfold]

I've been following this thread and decided to try my take in testing the Gastrus for bacteria. I crushed the tablet into powder and did a test in 4 ounces of whole milk @ 37 degrees Celsius in a glass jar with a cloth covering the top (for the carbon dioxide to be released) for 20 hours. My main point in testing was to make sure there was live bacteria. I had a decent result. Top of liquid was soft, almost like yogurt and middle had inoculate (cloudy mixture). I did a quick taste test and it tasted like unflavored Kefir. I am thinking of doing another test and boil the milk first (as I read earlier). Should I use raw milk or just the same store bought whole milk I used the first time? Any suggestions? Also, I put the jar in the fridge and the next day it is quite sour - much more than the day before. Btw, very interesting thread!

 

I was planning on doing this simple experiment as well. It looks like you've got a got a good result which is promising.

 

I'm waiting for my Gastrus to arrive and then I'll start doing the same. It would be interesting to hear if you feel any differences after consuming the fermented milk regularly.

[recon]

 

 

 

If it does contain the strain this would be patent infringement. You would have to take their word they expressed in a poorly made video. Why are none of the strains used in this supplement listed on the label? I would never consider buying this supplement.

 

 

Yeah I'm also dubious about it, I've sent them an email requesting more information and am awaiting a response. Will update when I hear back from them.

Is there any supplement containing just ATCC 6475?
Even Gastrus contains another Reuteri, I believe. Is there not one on he market just having that one particular strain?

 

Not that anyone knows of. Infact, the only one I'm aware of that definitely contains 6475 at all is the Gastrus product.

 

I wonder what's the rationale for that? I'm going to shoot them an email to ask.

[mkmicro]

As most of you, I have been trying to find ATCC 6475 strain but only find it in Gastrus. I have reached out to Lifeway that makes Kefir with Reuteri on what strain they use. Their response is "proprietary" and would not let me know what strain it was. I think it's more for protecting themselves from any health claims.

 

My second test:

 

I used raw cow milk (8 ounces). Boiled for 5 minutes.

Poured in glass jar and added broken up chunks of Gastrus tablet.

Put in warmer @ 95-100F overnight

Checked after 10 hours and found that the broken up chunks were still on the bottom starting to gel (at least it looked like gel). Decided to stir it to disperse the remnant chunks (I think crushing is best due to this find).

Next morning yogurt thickness with a little clumpy/runny on bottom

 

I think the test was successful for the most part. Should have left it in another 3-4 hours.

 

Taste is good - much better than store bought milk.

 

I want to try this on a bigger batch - a quart size. I'll use two tablets (crushed) in the same method as last test using raw milk. I'm thinking of buying a yogurt maker. Can the temp be controlled or is it set at a certain temp?

 

 

[mkmicro]

I've been doing research and talking with a friend who knows biology/chemistry. I keep getting asked why use milk - and SDXL brought this up as well. Anyone try a different method? Could this grow in a Kombucha mixture with extra glucose? Also, I've been taking the tablets and find it seems to be effective. Meaning, the day after I took the tablet my bowel movement was different. Seems like I have more and much larger. Been about a week and still the same. I'm thinking, should I even try to increase the strain? It is pricey but it seems to be working at 200M.

[sdxl]

In the past I have done something with microbiology. Knowing about stuff like growth curves, substrates, metabolites, buffers, pH control, osmotic stress, etc. could be useful if you want to grow bacteria. Go read a couple of microbiology books, read some relevant studies and patents if you really want to grow these bacteria. I may post some of my experiments in the future.

 

In a clinical trial they used 10 billion CFU daily of ATCC PTA 6475. If 100 million CFU would be sufficient, why would they give the women in the study daily 100 as much as in a Gastrus tablet? Seems to me that it requires more CFU to sort a significant systemic effect. Gastrus is meant as an adjunct treatment for those with H. pylori infections, targeting the GI tract. The reason why DSM 17938 is included in Gastrus is likely because it produces significantly more reuterin than ATCC PTA 6475.

 

The estimated study completion date is next month. The press release looks positive. Hopefully BioGaia considers to launch a single strain probiotic with ATCC PTA 6475 soon. If the product is lauched it will be likely a powder, just like in the clinical trial. BioGaia registered a new trademark this year, PROVESI. That could be it or it could be nothing relevant. Only time will tell.

 

[Izan]

 

In the past I have done something with microbiology. Knowing about stuff like growth curves, substrates, metabolites, buffers, pH control, osmotic stress, etc. could be useful if you want to grow bacteria. Go read a couple of microbiology books, read some relevant studies and patents if you really want to grow these bacteria. I may post some of my experiments in the future.

 

In a clinical trial they used 10 billion CFU daily of ATCC PTA 6475. If 100 million CFU would be sufficient, why would they give the women in the study daily 100 as much as in a Gastrus tablet? Seems to me that it requires more CFU to sort a significant systemic effect. Gastrus is meant as an adjunct treatment for those with H. pylori infections, targeting the GI tract. The reason why DSM 17938 is included in Gastrus is likely because it produces significantly more reuterin than ATCC PTA 6475.

 

The estimated study completion date is next month. The press release looks positive. Hopefully BioGaia considers to launch a single strain probiotic with ATCC PTA 6475 soon. If the product is lauched it will be likely a powder, just like in the clinical trial. BioGaia registered a new trademark this year, PROVESI. That could be it or it could be nothing relevant. Only time will tell.

 

that would be awesome! an atccc pta 6475 ONLY product in powder form!