Bacteria in a bottle, Myth or Fact

[brandon429]

Context matters


findings at a ph of 7 have no comparison to anything in reefing


that entire post above is for freshwater tanks where ammonia toxicity was never in debate here and has zero application here in this forum at all. I don’t even believe that author meant it for reefing I think you guys were having a freshwater tank discussion on Facebook and that was associated here since it was a bottle bac study

David isn’t wrong if you guys‘ discussion is kept in the exact context discussion (freshwater) you were having at the time.


ps: I wouldn’t have known the context of pH matters like it does without being informed by Dan so it’s not an intended distraction I do believe. In 2015 I’d have thought the same thing.

David isn’t wrong, it just wasn’t a reefing context discussion you were having and if he was informed of that application I’m sure he’d agree about the very different needs in saltwater tanks.

 

[Dr. Reef Verified Sponsor]

pH and Alk matters in saltwater for bacteria to thrive.
I did experiment on a turbostart 9 (refrigerator verion) to sit outside in my garage which normally is 100-120F and it kept working for 2.5 months before quit cycling tanks fast.

I have been using these bottles (all brands) and have not had one fail me. They are intended to work with dosing tank and adding fish, unlike i did where i had ammonia in tank at certain level and let bacteria bring it down,

Page 1 post 2and 3 shows all the winners in all the experiments i did.

 

[zeropauper]

Context matters


findings at a ph of 7 have no comparison to anything in reefing


that entire post above is for freshwater tanks where ammonia toxicity was never in debate here and has zero application here in this forum at all. I don’t even believe that author meant it for reefing I think you guys were having a freshwater tank discussion on Facebook and that was associated here since it was a bottle bac study

David isn’t wrong if you guys‘ discussion is kept in the exact context discussion (freshwater) you were having at the time.


ps: I wouldn’t have known the context of pH matters like it does without being informed by Dan so it’s not an intended distraction I do believe. In 2015 I’d have thought the same thing.

David isn’t wrong, it just wasn’t a reefing context discussion you were having and if he was informed of that application I’m sure he’d agree about the very different needs in saltwater tanks.

pH and Alk matters in saltwater for bacteria to thrive.
I did experiment on a turbostart 9 (refrigerator verion) to sit outside in my garage which normally is 100-120F and it kept working for 2.5 months before quit cycling tanks fast.

I have been using these bottles (all brands) and have not had one fail me. They are intended to work with dosing tank and adding fish, unlike i did where i had ammonia in tank at certain level and let bacteria bring it down,

Page 1 post 2and 3 shows all the winners in all the experiments i did.

Hi Brandon and Dr. Reef, I apologize if I was unclear about what I was asking.

I should have narrowed down to reddit thread to only the part where I was talking about the bottled bac experiment that David did. You can read about his methodology and conclusions here: https://web.archive.org/web/2021091...ndex.php/2-8-1-bacteria-in-a-bottle-in-depth/

Although his experiment was done in freshwater, I believe that the aims of both your experiment and his are similar - to determine if bottled bacteria do indeed quicken the cycling of a new tank.

I have seen your results on this thread that show how the different brands line up against one another, but I would like to ask if there was a control (no bottled bacteria) in your experiment to show that bottled bacteria indeed cycle tanks faster than without. I ask this because David on his website has made the claim that "absolutely NO tests by anyone with any data which proves the author wrong." So basically, he is saying that his results are irrefutable and bottled bacteria is a scam, which is the opposite conclusion from the experiments in this thread.

I've read your methodology and I fully understand why the parameters are as such. They are simple and easy to follow and can produce accurate conclusions.

But I can't say the same for David's experiment on his website. Here are the problems I've identified that might conflate the results of his experiment and render them inaccurate:

1) The main aim of his experiment was to determine if bottled bacteria cycled tanks faster. If that's the case, that should be the only changed variable. However, he added another variable (presence of phosphates).

2) David says that 3 experiments were done. 2 of which were with phosphate added and 1 without. And he also mentioned "seven of these (bottled bacteria) were replicated twice for accuracy".
- We all know the importance of experimental replication (especially in such a simple experiment even though it is of biological origin). If David wanted to do a phosphate experiment, it should've been a separate one conducted 3 times. And he should've replicated the non-phosphate experiment 3 times.
- Looking at his results table, I can't find the 7 bottled bacteria that were "replicated twice for accuracy". I count 9 that were replicated twice in the phosphate experiments.

3) If you read even further, the methodology suggests that even experiment 1 and 2 (with phosphate) was different. He changed 5 sources of bacteria for experiment 2.

4) Ignoring the fact that we only have 1 set of results without phosphate to look at, even the 2 controls are significantly different from each other (36 vs 52 days).

5) David claims that "Each bacteria-in-a-bottle product was used according to the directions on the respective container.", but declined to comment on whether he repeated the dosage on the following days in accordance to manufacturer's instructions (e.g. stability requires a larger dose on the first day followed by half dose daily for the next 7 days).

 

[MnFish1]

Hi Brandon and Dr. Reef, I apologize if I was unclear about what I was asking.

I should have narrowed down to reddit thread to only the part where I was talking about the bottled bac experiment that David did. You can read about his methodology and conclusions here: https://web.archive.org/web/2021091...ndex.php/2-8-1-bacteria-in-a-bottle-in-depth/

Although his experiment was done in freshwater, I believe that the aims of both your experiment and his are similar - to determine if bottled bacteria do indeed quicken the cycling of a new tank.

I have seen your results on this thread that show how the different brands line up against one another, but I would like to ask if there was a control (no bottled bacteria) in your experiment to show that bottled bacteria indeed cycle tanks faster than without. I ask this because David on his website has made the claim that "absolutely NO tests by anyone with any data which proves the author wrong." So basically, he is saying that his results are irrefutable and bottled bacteria is a scam, which is the opposite conclusion from the experiments in this thread.

I've read your methodology and I fully understand why the parameters are as such. They are simple and easy to follow and can produce accurate conclusions.

But I can't say the same for David's experiment on his website. Here are the problems I've identified that might conflate the results of his experiment and render them inaccurate:

1) The main aim of his experiment was to determine if bottled bacteria cycled tanks faster. If that's the case, that should be the only changed variable. However, he added another variable (presence of phosphates).

2) David says that 3 experiments were done. 2 of which were with phosphate added and 1 without. And he also mentioned "seven of these (bottled bacteria) were replicated twice for accuracy".
- We all know the importance of experimental replication (especially in such a simple experiment even though it is of biological origin). If David wanted to do a phosphate experiment, it should've been a separate one conducted 3 times. And he should've replicated the non-phosphate experiment 3 times.
- Looking at his results table, I can't find the 7 bottled bacteria that were "replicated twice for accuracy". I count 9 that were replicated twice in the phosphate experiments.

3) If you read even further, the methodology suggests that even experiment 1 and 2 (with phosphate) was different. He changed 5 sources of bacteria for experiment 2.

4) Ignoring the fact that we only have 1 set of results without phosphate to look at, even the 2 controls are significantly different from each other (36 vs 52 days).

5) David claims that "Each bacteria-in-a-bottle product was used according to the directions on the respective container.", but declined to comment on whether he repeated the dosage on the following days in accordance to manufacturer's instructions (e.g. stability requires a larger dose on the first day followed by half dose daily for the next 7 days).

the problem is he did not define what he meant by 'cycled'. But I think for the most part, you answered your own question the replicates are not 'the same'.

The comparators (positive controls) he used - like manure, and dry black soil likely have much higher bacterial concentrations than a bottled product. So - you really can't compare them well either.

The experiment here was designed to look at ammonia - and whether bacteria (bottled) affected it. So - you really can't compare the studies.

Its clear that bacteria need a phosphate source. of some kind.

 

[zeropauper]

The comparators (positive controls) he used - like manure, and dry black soil likely have much higher bacterial concentrations than a bottled product. So - you really can't compare them well either.

Right, I totally forgot about the point about manure and soil bacterial concentrations. I touched on that in my reddit thread.

How did he ensure that the amount of manure, soil or filter gunk was comparable to the amount of beneficial bacteria in commercial products? I can add a heap of soil that fills up half the 5 gallon bucket and it would obviously cycle much quicker because way more bacteria is present from the start.

The experiment here was designed to look at ammonia - and whether bacteria (bottled) affected it. So - you really can't compare the studies.

Actually, on page 56 I found that the results included a control, so even though it wasn't explicitly stated on the first page, the presence of a control allows both studies to be compared. That's also the reason why I asked Dr Reef if the experiments had controls.

 

[brandon429]

Here’s another way to confirm with a simple control

get a seneye nh3 meter do *not* use colorimetric test tube kits for this proof they’re too slow to register and inaccurate

get seneye or dont do it / accept Dr. Reef’s findings without hesitation until a seneye is procured to disprove the need for bacterial support in reef tank bioload carry

-set up the seneye and benchmark the slide for it and the probe on a fully running reef tank with corals and fish. A matured one, a full running reef tank. The meter will read .002-.009 nh3 or it’s not trimmed/ something is wrong with the slide prep or handling if it doesn’t, try a new slide and pre soak them 48 hours or however long the directions say to prep them. Once you have a seneye that reads .002-.009 on a running reef tank that’s calibrated and ready. This is the nh3 range display tanks run at

move the sensor over to a five gallon bucket insta reef half full with dry Marco rocks and water set to reef salinity and heated/ circulated


put in two clownfish and give them a few feed pellets

as soon as that meter hits .01 pull them out before they die, that’s proof you need bacteria in saltwater and that these statements above have zero basis in reefing and are solely relayed from the realm of freshwater and can’t be even haphazardly applied here. No bottle bac will permit this rise, no cycled display will permit an overnite rise to .01 and that’s what will happen in the control.

 

[brandon429]

zeropauper if you could recruit folks off Facebook or local reefing circles for some critical data it’d be very appreciated here

see what MSteven1 did on page 97 onward. That’s a very very uninvestigated realm within cycling circles, the fabled and as you can see highly debated fully unassisted marine self cycle. It’s what David is claiming to happen rather quickly in freshwater and I agree it is pretty quick. A common cycling chart shows 30 days to ready across all books, across all websites for that reason of inherent consistency and sourcing from nature


nobody can remark on such a chart for marine biofilter self establishment and MSteven’s work is the first I’ve seen. When attaining this simple time-based experiment take extra care not to contaminate or vector in bacteria from any running tanks, you can see that’s the doubt given to Stevens documentation on page 97+ so take care not to use parts in the setup that could bring in marine- selected bacteria. The final proofing of ability will need to be done with liquid ammonium chloride as the test loading, using any animals in this experiment is an instant bacterial vector


if you could source out some easy proofing for marine unassisted cycles that’s appreciated and very very relevant to needed data for the hobby

 

[Dr. Reef Verified Sponsor]

Right, I totally forgot about the point about manure and soil bacterial concentrations. I touched on that in my reddit thread.




Actually, on page 56 I found that the results included a control, so even though it wasn't explicitly stated on the first page, the presence of a control allows both studies to be compared. That's also the reason why I asked Dr Reef if the experiments had controls.

There was a control tank in every study, Control had no bacteria added and it cycled the last in every study,

In conclusion,
A 5 gal Tank cycled from 2-8ppm fastest with Fritz turbostart 900 around 3-5 days. (no help or food was needed, did cycle faster if food was present) (food adds carbon source)
2nd best were Bio Spira and Dr Tims one and only cycled 5 gal from 2-8ppm ammonia to 0 in 5-7 days. (helped cycle faster with food)
Rest all products stalled without carbon source (food or live fish) so i had to add food and within a day or 2 of adding food stalled ammonia went to 0.

Thus again my experiment was a bit flawed in a sence that these products are not to be used to bring ammonia down but rather dont let ammonia creep up on a fish in a non cycled tank.

Now a days like i mentioned before I am in business of selling quarantined fish, If something goes wrong in one of my tanks, I add bleach to running tank and few hours later drain and dry, after 24 hrs of drying I put new sand and ceramic media and fill with saltwater and add turbostart900 and add few small or med fish. Never lost one to ammonia/not cycled tank.
I do this tear down and resetup almost weekly basis due to large amounts of fish i qt and number of tanks i have.

Also these bacteria in bottle do well when you cycle a less salinity water like 1.017-19 with alk and pH high. Add food or live fish and dose bacteria. Dont add a ton of fish more like 3-4 small fish or 1-2 med and then let it build up.

 

[Dr. Reef Verified Sponsor]

Another thing regarding ammonia is that there is still alittle play (not much) but just a little.
There are 2 types of ammonia in the tank. NH3 which is in gas form, deadly to fish and NH4 the non toxic kind.
If the pH was to be between 8.2-8.4 dosing ammonia or leaving a deli shrimp in tank will produce both kinds of ammonia in tank which would be in a specific ratio. Say you tested 2 ppm on your test kit which every hobby level kit on market is a TAN kit (total ammonia NH3/NH4) They dont break the toxic from non toxic apart. Only kit i am aware of that does both apart is Seachem multitest ammonia kit.
So if you had 2 ppm on a TAN kit its safe to assume toxic ammonia NH3 is about 0.2 ppm.

Seneye unit monitors the deadly type of ammonia NH3.

NH3 chart (according to Seneye)

Safe from 0.001 to 0.02
Alert from 0.02 to 0.05
Alarm from 0.05 to 0.2
Toxic from 0.2 to 0.5
Deadly 0.5+

Thus if you are using a TAN kit asuming pH and other prameters are all in check a reading of 1ppm is equal to 0.1 which is alarming but not deadly, 0.5ppm is equal to 0.05 which is on the alrt side again not deadly.
So like most people report 0.25ppm reading on most ammonia tests, there is not much to be alarmed on, as NH3 will be in safe zone and not deadly.

 

[MnFish1]

Right, I totally forgot about the point about manure and soil bacterial concentrations. I touched on that in my reddit thread.




Actually, on page 56 I found that the results included a control, so even though it wasn't explicitly stated on the first page, the presence of a control allows both studies to be compared. That's also the reason why I asked Dr Reef if the experiments had controls.

Yes - I meant to mentioned - there was a control!

 

[mcgullen]

Resurrecting this thread bc it's an amazing read.

 

[mcgullen]

So I have been paying for heterotrophic sludge removers :(

 

[MnFish1]

So I have been paying for heterotrophic sludge removers :(

Curious - are they helping? If so they are worth your money:). The main focus of this thread was the bacteria used for cycling - as comparing to weekly or regular doses of bacteria. FWIW - heterotrophic bacteria also remove ammonia - and can be used for cycling. The goal was to try to show whether bacteria that only use ammonia can exist in a bottle (many thought it was not possible).

 

[NeoAqua]

I have a naive question. Will all these bottled bacteria workin RO + UV water or DI water? Won’t that be the best proof whether they work or not. Because RO + UV or DI water will not have any bacteria of any kind.

On a side note before I found this amazing thread, I bought fish and introduced them on day 1 with Seachem Stability. As on day 7 ammonia reads between 0.5ppm to1ppm. It has not reduced. No I do partial water change everyday.

 

[vetteguy53081]

I have a naive question. Will all these bottled bacteria workin RO + UV water or DI water? Won’t that be the best proof whether they work or not. Because RO + UV or DI water will not have any bacteria of any kind.

On a side note before I found this amazing thread, I bought fish and introduced them on day 1 with Seachem Stability. As on day 7 ammonia reads between 0.5ppm to1ppm. It has not reduced. No I do partial water change everyday.

How are you testing ammonia which may be false readings?
Bacteria is beneficial for building denitrators and not an immediate ammonia/nitrate reduction.
Take a water sample to an LFS that does not use API tests and see what readings they get and to compare with yours.
Doing 2 gallon water changes for two weeks will reduce these numbers

 

[Lasse]

Will all these bottled bacteria workin RO + UV water or DI water? Won’t that be the best proof whether they work or not. Because RO + UV or DI water will not have any bacteria of any kind.

It will probably kill them because of osmosis. Salt out from the bacteria - water in. RODI may have encapsulated bacteria or spores that can stand the forces of osmosis.

Sincerely Lasse

 

[MnFish1]

I have a naive question. Will all these bottled bacteria workin RO + UV water or DI water? Won’t that be the best proof whether they work or not. Because RO + UV or DI water will not have any bacteria of any kind.

No. But first why would you put bottled bacteria in RO/DI + UV water?

On a side note before I found this amazing thread, I bought fish and introduced them on day 1 with Seachem Stability. As on day 7 ammonia reads between 0.5ppm to1ppm. It has not reduced. No I do partial water change everyday.

And you re-added the stability each day? the ammonia (if accurate - and I would double check your tests) is really a little too high for healthy fish. Are you sure you might not have overdone it - with regards to the fish you added?