Prodibio BioDigest Likely Has No Effect On Ammonia Oxidation and Does Not Help with Waste Reduction

[mcgullen]

No positive results whatsoever.

I have repeated all model tank tests with same results.

I have used BioDigest as instructed on established aquariums and monitored their TAN, PO4 and NO3 levels, only to see no difference as compared to controls for 2 weeks.

Is there anyone who knows what BioDigest is actually supposed to do and how to test for such results?

 

[ReefLife_Guy]

No positive results whatsoever.

I have repeated all model tank tests with same results.

I have used BioDigest as instructed on established aquariums and monitored their TAN, PO4 and NO3 levels, only to see no difference as compared to controls for 2 weeks.

Is there anyone who knows what BioDigest is actually supposed to do and how to test for such results?

You shouldn’t trust any company’s proprietary blend of anything without supporting evidence that it does what they claim. It is not on the consumer to do the experiments to validate a companies product. I don’t even see an official website for this product. There is no FDA regulations for these products, they could literally sell you water in a bottle and say it will do anything and have no need to back up what they say.

I think this hobby is at a point where we should force companies to provide the evidence that their product works. Unless the technology is obviously well validated for the uses they are using them for, in which case I would only care to see quality control measures for the product.

For example, the technology that aquabiomics uses to analyze your tank’s “microbiome” is a well known, established, and valid method for sequencing-based bacterial analysis. The only thing I would expect as a consumer is some evidence that they are providing high quality data with appropriate QC and standards used. If they claimed using their product would do x, y, and z for your aquarium, well then at that point I would expect to see some preliminary data at minimum.

At this point I think if the company cannot provide simple evidence to support their claims (whether or not that evidence is their own) then you should just spend your money elsewhere.

 

[taricha]

Is there anyone who knows what BioDigest is actually supposed to do and how to test for such results?

So far, tests that simulate real world scenarios all failed to elicit beneficial responses clearly attributable to BioDigest.

I totally understand if you decide that under the scenarios you envision (established tanks) - it doesn't work for things people expect, and you'd like to move on.

But just because I think it's interesting let me point out a particular scenario/question that I think is worth talking about.
Let's talk about the case for prodibio as a cycling / tank starter bacteria.

2. Prodibio BioDigest Likely Doesn't Help with Ammonia Oxidation

One would have hoped that, even if BioDigest does not contain true nitrifying bacteria, at least its heterotrophic strains could still consume ammonia when rapidly multiplies.

Well, it didn't.

Prodibio BioDigest did not bring down ammonia in any of the three runs within 48 hours. Nor did BioDigest beat the control tank in any of the run. In the second run after 72 hours, both the control tank and the BioDigest tank became cloudy, with ammonia starting to drop. This suggests that external heterotrophic bacteria started to multiply and brought ammonia down.

But, as you note - in the bottle bacteria myth or fact thread Dr Reef found that a number of bacteria did not consume any ammonia until some fish food was added - then all of the previous non-responders did in fact consume ammonia. This was interpreted as these types being heterotrophic nitrifiers - needing some amount of carbon in order to process ammonia.
So there's a couple of interesting (to me, anyway) possibilities I can think of.
One is what you envision below: it was actually bacteria already present in the new cycling tanks that processed the ammonia, and maybe not clear that the bottled bacteria were needed, heterotrophs capable of doing this might be everywhere anyway.

I suspect this is what happened in @Dr. Reef study. When he added fish food and witnessed ammonia starting to drop in the BioDigest tank, it was probably some naturally occurring Bacillus strains that were already in the tank water doing the job.

This is plausible. I could not find in the Myth or Fact thread where the result with heterotroph nitrifiers was done using a no-bacteria-product control. So those other products may not have done it either. Could just be heterotroph contamination consuming the ammonia.

Another possibility is that the Myth or Fact thread conditions weren't quite replicated here. Namely, I think in that thread the tanks were bleached and restarted with new water, so the bottled bacteria had a better chance to show their stuff. I'm under the impression that you used tank water as the water you added prodibio to. So that wouldn't be quite the situation for a new dry rock start.


I wonder if you more tightly replicated the Myth or Fact setup and bleached/washed containers with new saltwater and dry sand and did ammonia + crushed fish food in tanks with 1) No added bacteria, vs 2) prodibio, vs 3) a popular tank starter heterotroph in separate containers, would the no bacteria actually do nothing to the ammonia (or are heterotropjh nitrifiers everywhere)? Would prodibio actually underperform other popular heterotroph tank starters?

 

[mcgullen]

You shouldn’t trust any company’s proprietary blend of anything without supporting evidence that it does what they claim. It is not on the consumer to do the experiments to validate a companies product. I don’t even see an official website for this product. There is no FDA regulations for these products, they could literally sell you water in a bottle and say it will do anything and have no need to back up what they say.

I think this hobby is at a point where we should force companies to provide the evidence that their product works. Unless the technology is obviously well validated for the uses they are using them for, in which case I would only care to see quality control measures for the product.

For example, the technology that aquabiomics uses to analyze your tank’s “microbiome” is a well known, established, and valid method for sequencing-based bacterial analysis. The only thing I would expect as a consumer is some evidence that they are providing high quality data with appropriate QC and standards used. If they claimed using their product would do x, y, and z for your aquarium, well then at that point I would expect to see some preliminary data at minimum.

At this point I think if the company cannot provide simple evidence to support their claims (whether or not that evidence is their own) then you should just spend your money elsewhere.

I am based in HK so don’t have easy access to Aquabiomics

I totally understand if you decide that under the scenarios you envision (established tanks) - it doesn't work for things people expect, and you'd like to move on.

But just because I think it's interesting let me point out a particular scenario/question that I think is worth talking about.
Let's talk about the case for prodibio as a cycling / tank starter bacteria.




But, as you note - in the bottle bacteria myth or fact thread Dr Reef found that a number of bacteria did not consume any ammonia until some fish food was added - then all of the previous non-responders did in fact consume ammonia. This was interpreted as these types being heterotrophic nitrifiers - needing some amount of carbon in order to process ammonia.
So there's a couple of interesting (to me, anyway) possibilities I can think of.
One is what you envision below: it was actually bacteria already present in the new cycling tanks that processed the ammonia, and maybe not clear that the bottled bacteria were needed, heterotrophs capable of doing this might be everywhere anyway.

This is plausible. I could not find in the Myth or Fact thread where the result with heterotroph nitrifiers was done using a no-bacteria-product control. So those other products may not have done it either. Could just be heterotroph contamination consuming the ammonia.

Another possibility is that the Myth or Fact thread conditions weren't quite replicated here. Namely, I think in that thread the tanks were bleached and restarted with new water, so the bottled bacteria had a better chance to show their stuff. I'm under the impression that you used tank water as the water you added prodibio to. So that wouldn't be quite the situation for a new dry rock start.


I wonder if you more tightly replicated the Myth or Fact setup and bleached/washed containers with new saltwater and dry sand and did ammonia + crushed fish food in tanks with 1) No added bacteria, vs 2) prodibio, vs 3) a popular tank starter heterotroph in separate containers, would the no bacteria actually do nothing to the ammonia (or are heterotropjh nitrifiers everywhere)? Would prodibio actually underperform other popular heterotroph tank starters?

Thanks for the suggestion!

I should point out that in my very first test, I tried to replicate what Dr Reef saw.

1. I used fresh RO water to simulate freshwater tanks BioDigest advertises as compatible with.

2. I did sterilize all tanks with ClO2 tablets. This was done for not just the first test but all subsequent tests. Also to remove the possibility of residual chlorine interfering with bacterial products, sterilized tanks were rinsed with RO water with Na2S2O3.

3. I used household brown sugar as carbon and not fish food in my first tests. Sucrose, ethanol and sodium acetate were used for subsequent freshwater and saltwater tests.

Ammonia never dropped in any test in BioDigest tanks with 0.5, 1, or 5ppm as initial concentration in a way that’s clearly attributable to BioDigest. Ammonia only dropped when both experiment and control tanks started to show bacterial bloom after 48 hours that were likely to be caused by bacterial source not related to BioDigest. In other words, experiment results do NOT support a claim that the concentrated strains in BioDigest would give experiment tanks a head start.

In light of the above, is there anything else that I should try?

Thanks.

 

[taricha]

Thanks for the suggestion!

I should point out that in my very first test, I tried to replicate what Dr Reef saw.

1. I used fresh RO water to simulate freshwater tanks BioDigest advertises as compatible with.

2. I did sterilize all tanks with ClO2 tablets. This was done for not just the first test but all subsequent tests. Also to remove the possibility of residual chlorine interfering with bacterial products, sterilized tanks were rinsed with RO water with Na2S2O3.

3. I used household brown sugar as carbon and not fish food in my first tests. Sucrose, ethanol and sodium acetate were used for subsequent freshwater and saltwater tests.

Ammonia never dropped in any test in BioDigest tanks with 0.5, 1, or 5ppm as initial concentration in a way that’s clearly attributable to BioDigest. Ammonia only dropped when both experiment and control tanks started to show bacterial bloom after 48 hours that were likely to be caused by bacterial source not related to BioDigest. In other words, experiment results do NOT support a claim that the concentrated strains in BioDigest would give experiment tanks a head start.

In light of the above, is there anything else that I should try?

Thanks.

Thanks. Very much appreciate the detail.
This discussion makes me wonder if there's a real possibility that several "instant cycle" heterotroph products may be benefitting from random contamination that is unavoidable, and their product may activate and simply participate along with the bacteria everpresent in the environment - not necessarily be the primary driver of the heterotrophic ammonia processing.
But back to prodibio...

a google scholar for pseudomonas in aquaculture is a good way to figure out what somebody might hope their bacterial product can do. Mostly the game these papers are playing is to find some strains that show up in high nutrient aquaculture water, and measure their abilities to lower ammonia, nitrite, nitrate when fed different amounts and sources of carbon. What makes them desirable or interesting is that presumably they can remove a higher amount of these N sources than random other bacteria given the same C/N ratio.

You found pseudomonas in the bottle, so lets presume that prodibio is not finding some super obscure never-before studied strain, and culturing it en masse. They are probably using some strain that is studied and has been demonstrated to culture up and be salt/fresh tolerant.

If I were really motivated to try to see the psuedomonas in prodibio do something that might be relevant, I'd set up a side by side with two replicates of new salt water, and two replicates of new saltwater with prodibio. I'd add 50 mg/L of crushed fish flake ( at ~50% protein = 4ppm Nitrogen) to each and aerate each of them and track the ammonia over a week.

In theory there should be differences in the trend of ammonia over time vs the two treatments - if the prodibio is doing something - with a lower stable ammonia level after one week.

 

[mcgullen]

Thanks. Very much appreciate the detail.
This discussion makes me wonder if there's a real possibility that several "instant cycle" heterotroph products may be benefitting from random contamination that is unavoidable, and their product may activate and simply participate along with the bacteria everpresent in the environment - not necessarily be the primary driver of the heterotrophic ammonia processing.
But back to prodibio...

a google scholar for pseudomonas in aquaculture is a good way to figure out what somebody might hope their bacterial product can do. Mostly the game these papers are playing is to find some strains that show up in high nutrient aquaculture water, and measure their abilities to lower ammonia, nitrite, nitrate when fed different amounts and sources of carbon. What makes them desirable or interesting is that presumably they can remove a higher amount of these N sources than random other bacteria given the same C/N ratio.

You found pseudomonas in the bottle, so lets presume that prodibio is not finding some super obscure never-before studied strain, and culturing it en masse. They are probably using some strain that is studied and has been demonstrated to culture up and be salt/fresh tolerant.

If I were really motivated to try to see the psuedomonas in prodibio do something that might be relevant, I'd set up a side by side with two replicates of new salt water, and two replicates of new saltwater with prodibio. I'd add 50 mg/L of crushed fish flake ( at ~50% protein = 4ppm Nitrogen) to each and aerate each of them and track the ammonia over a week.

In theory there should be differences in the trend of ammonia over time vs the two treatments - if the prodibio is doing something - with a lower stable ammonia level after one week.

The control tanks showed bacterial bloom hours before Prodibio tank did on day 3 and control tanks’ ammonia level dropped rapidly..

 

[Laith]

Interesting discussion!

I started using Biodigest in my previous 250l reef tank but I always dosed Bioptim along with it (as recommended on their website), every two weeks. I did notice an improvement, not so much in phosphate reduction but it did stabilize NO3 that was rising at a steady level. I also noticed that my sand bed seemed much cleaner but this is always a subjective view.

But one of my defaults is I tend to try several things simultaneously so I'm never 100% sure what action gave the result that I saw.

I set up my current 1900 liter reef in October of 2019 and decided to continue using the combination of Biodigest and Bioptim every two weeks since then.

Two questions:

1. The OP decided not to use Bioptim in the tests (as far as I understood). If that is the case, maybe redo the tests using the Bioptim as well? As far as I can tell, Bioptim is some type of food (carbon?) for the bacteria plus some other elements.

2. If it can be shown that Biodigest (with Bioptim) has absolutely no effect on a reef tank then I would of course seriously think about stopping using them (they are not cheap, especially on a big tank). However, here is a scenario that I would like to avoid:

I have been dosing the two products for three and a half years on a regular basis. I would imagine that the products are adding SOMETHING to the tank. I am worried that by suddenly stopping, some type of biological equilibrium would be disturbed and I could face multiple other problems...

Has anyone stopped using these products from one day to the next after long term use? If so, what were the consequences, both good or bad?

Thanks!

 

[Mattiejay6]

@Reefahholic interesting read here!

 

[Reefahholic]

But what I can say is that BioDigest does not appear to contain any nitrifying bacteria in the strict sense.

I could see that being the case. Nitrifying bacteria has little effect on nutrient levels when initially dosed. Heterotrophs on the other hand have a major impact, compete for surface area and nutrients, divide in 20 minutes, and can rapidly dominate the Nitrifiers. They just do not grow as fast. Nutrients being impacted in 1-2 days has been reported with Biodigest, so I think it’s mostly or all Heterotrophic bacteria. I don’t prefer to dose an entire vial to my system at once as I don’t like “big impacts” so I choose to dose products where I can dose very small amounts more frequently and have more control. I mean I guess I could break the vial and store it in something to reduce the amount dosed, but it’s too much trouble.

 

[Reefahholic]

I have used BioDigest as instructed on established aquariums and monitored their TAN, PO4 and NO3 levels, only to see no difference as compared to controls for 2 weeks.

What all was being dosed to the tank? Feeds, etc…

 

[Reefahholic]

I’ve dosed an entire bottle of “One and Only” that has pure nitrifiers, and the impact on nutrients is basically nonexistent or takes place at a much slower rate.

On the flip side if you dose an entire bottle of Waste Away you would nuke the entire tank. Go read the reviews at BRS.

I always tell people to start with 1/8 of the recommended dose.

 

[mmoner]

Saw this in an insta post.

 

[taricha]

Saw this in an insta post.

is that purported to be a sample of the product? Because that's just a snapshot of what a reef tank microbiome would look like.
Highly skeptical any sealed vial is delivering all those families live.

 

[taricha]

I don't like my first interpretation of the results....

Because that's just a snapshot of what a reef tank microbiome would look like.
Highly skeptical any sealed vial is delivering all those families live.

Saw this in an insta post.

Poked around a bit more and I think that the source of this is Jim Graham "telegraham", electrical engineering background who does a bunch of testing of electrical stuff and I think he's fully capable of correctly sampling the contents of the vial correctly for this microbiome test.

A better way to think about this result is "what is provided in big doses here that ISN'T in tank water?" and could be thought of as additions from this product.

so if we compare a typical biome result ...

...to what tests showed on the prodibio vial....

Then we'd say the major contributions from the product are Aeromonadaceae and Pseudomonadaceae.


This agrees with the original post in this thread...

The 16s rRNA results do support this claim as there is a Pseudomonas strain known to be an aerobic denitrifican present in the vial in high concentration.

The other stuff is a lot of families in typical reef tank samples. Maybe small amounts of a few other things.

 

[mmoner]

Do not know about him but nowadays Aquabiomics tests should answer whats in the bottle or not ? May be we need a guide what strains do the most work and how?

 

[Randy Holmes-Farley]

Do not know about him but nowadays Aquabiomics tests should answer whats in the bottle or not ? May be we need a guide what strains do the most work and how?

That’s the main issue with such testing, IMO. There is little known to establish what bacteria do what under what circumstances in a typical reef tank.

 

[ReefLife_Guy]

I might think about this from a different perspective because I studied host microbe interactions in grad school and currently collaborate on studies with microbiome sub studies but I don't think the greatest utility of this testing is for a single snapshot of a result. I think the best way to think about how to utilize this data is how we think about it as a methodology in scientific investigation. You use these kinds of shotgun metagenomic methodologies to assess patterns and to generate hypotheses but would never publish a conclusion based on this data without follow-up tests. While those follow-up tests might not be available for the hobby, you can still have utility in these tests if you have capability of interpreting the results with the understanding what limitations exist.

In my opinion this technology is too complex for lay people to interpret themselves. There is so much nuance with interpretation that is not general knowledge. The typical hobbyist don't know things like that there are strain level differences between bacteria in the same species that are so different it means the difference between a pathogenic organism and a beneficial organism. I am not aware that these results are provided in such detail but someone's sample that is positive for serratia marscescens could be the PDR60 strain that was identified as a coral pathogen for elkhorn acropora or it could be the CL1502 strain identified in some north atlantic sediment that has capabilities of heterotrophic nitrification and aerobic dentrification which most would agree they want in their tank.

It is the same reason doctors discuss results with their patients in the context of the patients situation taking into account everything they know at the time and the limitations of the test and don't just let them interpret their own results because........ biology is nuanced.

I went a little off tangent because I'm passionate about this topic but I think this technology has really changed science and medicine for the better but it is not without limitation. I think we have established that there is some level of diversity that correlates with a healthy tank but that is not to say there are not thriving tanks with low diversity. But it could also be true that lower diversity means a tank has lower resilience and those thriving tanks just haven't been stressed enough to show that. Where I think this technology is most helpful is comparing baseline relative abundance of populations when your tank is doing well compared to when your tank is not doing well and what populations have changed the most.

It is also a method for generating hypotheses when things aren't going right in your tank. Even if the results can't pinpoint a problem I think the more data I have to generate a probable causality of my issues the better. It also provides you the opportunity to test whether your interventions are working towards the change you wanted to make whether that was right or not. I think we can learn a lot in the hobby from this approach. I think publishing de-identified datasets along with hobbyist-reported tank values, conditions, inhabitants, etc into some kind of publicly available database would also be EXTREMELY helpful for us to investigate patterns and learn from each others data. With enough data we could answer or at least get a lot closer to an answer for a lot of these questions we have about reef tank microbiomes.

 

[mmoner]

Thank you...May be the tanks covered in such studies also should be classified with the bacteria products used to startup or maintained with. This also could be a clue if the product really helped (or not). In my local area, we have seen some tanks which started with Fritz9 had struggled getting their NO3-PO4 levels up (to minimums) for a long time so they could easily go for Pappone.

 

[BZOFIQ]

Subscribed.

 

[ReefLife_Guy]

Thank you...May be the tanks covered in such studies also should be classified with the bacteria products used to startup or maintained with. This also could be a clue if the product really helped (or not). In my local area, we have seen some tanks which started with Fritz9 had struggled getting their NO3-PO4 levels up (to minimums) for a long time so they could easily go for Pappone.

Agree the more data the better. Of course this database would have its own set of limitations when we are talking about the collection of non-standardized data points but you would be surprised how many revelations and great hypotheses come from less than ideal datasets. If you look at clinical trials there are a lot of preliminary trials that rely on positive signals from what we call patient reported outcomes. Which are basically just the patients telling us how they feel their symptoms are during a clinical trial which sounds extremely subjective but if the effect size of the intervention is strong enough, you will clearly pick up on a pattern despite all the confounders and other variables that can't be controlled for.

I have not yet used aquabiomics although I am curious and will sometime in the near future, but I believe they do collect some basic information from you to help them with interpreting your data. One might think it wouldn't be useful to collect this information to publish alongside the microbiome data, knowing that one hobbyist might report phosphates from their ICP test while another is relying on a slight difference in color with an at home test but when you get enough of these data points these kinds of variabilities and inaccuracies from self-reported data cannot mask really strong correlations. There is so much data that we get from these kinds of next-generation sequencing platforms that funding organizations like NIH have basically forced people to make their datasets publicly available because from a single good experiment you could have 20 different investigators make whole careers out of following up on different aspects of the data because the amount of data these output is wild.

I think the naysayers of what aquabiomics is doing, either don't understand the technology and what it can and cannot do or have unrealistic expectations of how complex biology is and don't want to hear nuanced answers to the issues they seek to resolve.