Looking for the Magic in Bottled Bacteria (and in our tanks)

[taricha]

@flampton is correct that there are no nitrifiers / tank cyclers doing anything important here. (Aside: in a flask of fish flake, aquarium water took 4.5-5 weeks to cycle)

But the nitrifiers aren't the ones we're talking about when we think about what happens to uneaten food, dead organisms, nameless grunge in our tank.
for @Variant 's point "Beneficial" is I suppose subject to interpretation.

Let me try to set goals for my bacteria
I'll tell you what I want to happen to food and waste that go in my tank - I want it to disappear totally! However, since that is impossible, I want what remains to be empty of nutrients that will drive nuisance growth. I'd like organic carbon to have already been consumed, and the same for nitrogen stores - so that nuisance algae are not fed by the particles that remain in the tank.

So if I have bacteria that consume nutrients C, N, P from fish food and digest it - either by converting it to their biomass or kicking it into the water, then I'm happy.

In that sense, my tiny doses of aquarium water are "beneficial" to my goals.

 

[taricha]

And as for the OPs flake food might have a preservative that would inhibit the bacterial growth. What was the ingredient list? Anything jump out at you taricha?

a later higher concentration (50mg/L protein) of same flake that gave nothing earlier gets the growth response we hope for. I'll post ingredients later. some flakes (not mine) have bacillus spores built right in!

edit: Ocean Nutrition Prime Reef Flakes

Ingredients
Plankton, dried fish protein digest, salmon, fish meal, wheat gluten, wheat flour, wheat starch, brine shrimp, lecithin, cochineal (added color), salmon eggs, fish oil, annatto seed (added color), paprika, dried kelp, kelp meal, mussels, choline chloride, magnesium sulfate, iron, inositol, a-tocopherol acetate (source of vitamin E), zinc sulfate, manganese sulfate, calcium carbonate, biotin, calcium pantothenate, niacin, sodium selenite, pyridoxine hydrochloride (source of vitamin B6), riboflavin (source of vitamin B2), L-ascorbic acid (source of vitamin C), potassium iodide, vitamin A supplement, citric acid (preservative), folic acid, thiamine mononitrate (source of vitamin B1), menadione sodium bisulfite complex (source of vitamin K3 activity), cobalt carbonate, vitamin B12 supplement.

Guaranteed Analysis
Crude Protein (min.) — 53.4%
Crude Fat (min.) — 16.6%
Crude Fiber (max.) — 0.2%
Moisture (max.) — 8.0%
Ash (max.) — 4.2%
Phosphorus (min.) — 0.6%

Sounds tasty!

 

[flampton]

@flampton is correct that there are no nitrifiers / tank cyclers doing anything important here. (Aside: in a flask of fish flake, aquarium water took 4.5-5 weeks to cycle)

But the nitrifiers aren't the ones we're talking about when we think about what happens to uneaten food, dead organisms, nameless grunge in our tank.
for @Variant 's point "Beneficial" is I suppose subject to interpretation.

Let me try to set goals for my bacteria
I'll tell you what I want to happen to food and waste that go in my tank - I want it to disappear totally! However, since that is impossible, I want what remains to be empty of nutrients that will drive nuisance growth. I'd like organic carbon to have already been consumed, and the same for nitrogen stores - so that nuisance algae are not fed by the particles that remain in the tank.

So if I have bacteria that consume nutrients C, N, P from fish food and digest it - either by converting it to their biomass or kicking it into the water, then I'm happy.

In that sense, my tiny doses of aquarium water are "beneficial" to my goals.

Now not sure what the end goal of your experiment is but if you want to actually start culturing your bacteria for some reason I would either A. Use bottled cultures as your starting point or B. You can use your aquarium bacteria but you would need to isolate individual strains. Then identify them somehow. The reason being is that you're culturing unknowns, and so if you get unlucky there's a chance a pathogen will be amplified, then you're going to stick that back in the aquarium at high concentration.

I would use A, haha. Once those cultures are established they'll do just as well as the aquarium bacteria.

 

[flampton]

Oh and adding bacteria to an already establish aquarium is a gimmick. The bacteria will grow with the available nutrients. So adding more isn't really going to do much. As far as what I'm interested in doing is isolating strains that have a high propensity towards polyphosphate storage. Then if you buy my product it would be one add and then done. I'm also looking into specialized carbons that only certain bacteria can consume. That way you can actually control carbon dosing to non pathogens. Again though it would be an add once product. So unfortunately I won't become a millionaire from forcing gimmicks on the average reefer.

 

[Dan_P]

@taricha Can you tell me if I'm interpreting your experiment incorrect?

So if you used aquarium water to see how it compares to bottled bacteria, would the results you're seeing prove that beneficial bacteria does indeed exist in the water column? Often times, I hear/read how beneficial bacteria is primarily on surfaces versus water borne. I recently watched the 2019 MACNA video of Dr.Tim's take on growing bacteria and he had stated how aquarium water doesn't have beneficial bacteria and that if you took water from a mature aquarium and dumped it in a new tank, you only just added dirty water.

Your results seem to show that the water itself has benefit. Am I wrong?

In what context did Dr. Tim define beneficial? Nitrate reduction? Organic carbon reduction?

Did Dr. Tim provide any data in support of the notion that aquarium water does not have beneficial bacteria? And beneficial to what goal?

 

[Nano sapiens]

Oh and adding bacteria to an already establish aquarium is a gimmick. The bacteria will grow with the available nutrients. So adding more isn't really going to do much. As far as what I'm interested in doing is isolating strains that have a high propensity towards polyphosphate storage. Then if you buy my product it would be one add and then done. I'm also looking into specialized carbons that only certain bacteria can consume. That way you can actually control carbon dosing to non pathogens. Again though it would be an add once product. So unfortunately I won't become a millionaire from forcing gimmicks on the average reefer.

While I try to dissuade new reefers from wasting their money on multiple infusions of 'Bottle Bac' for the same reef system whenever I see the question raised, it's hard to compete with commercial marketing interests.

Integrity seems to be an increasingly rare commodity these days. Best of luck in your endeavor!

 

[flampton]

While I try to dissuade new reefers from wasting their money on multiple infusions of 'Bottle Bac' for the same reef system whenever I see the question raised, it's hard to compete with commercial marketing interests.

Integrity seems to be an increasingly rare commodity these days. Best of luck in your endeavor!

It ain't just new reefers You got the probiotic method, lol

 

[Nano sapiens]

It ain't just new reefers You got the probiotic method, lol

All we are missing is the 'prebiotics'

 

[CoralClasher]

I’m hoping this discussion leads to a “probiotic”.

 

[Dan_P]

Conclusion of Part 1: Waking up Sleeping Beauty (for real this time)

When we left off in post 1 we had tried fish flake and skimmate, and our bottled bacteria products did precisely nothing. Not consuming PO4 to build biomass, not consuming Oxygen (oxidizing Carbon for energy), and not breaking down proteins etc to release N, seemingly not waking up.
Makes us worried if there are viable bacteria in the bottles. Let's go way out of the box and try something that isn't an aquarium thing at all, but a common bacterial growth medium - LB media.

Tank water was spiked to 1% LB media, heat-killed and added treatments of killed Waste Away "WA(st)", raw Waste Away "WA", or Aquarium water "Aq"
150mL flasks (day 3 measurements in extra 20mL test tubes). "heat-killed" media and Waste Away were done by covered 30 min boiling in tightly covered pot. In order to allow more growth with less restriction from O2 in this rich media, 10% air space was left in each sample, so O2 could diffuse in slowly.

And Voila! we have vigorous activity. By day 5 we have complete O2 consumption by the bottled (and aquarium) bacteria and plenty of Ammonia production. Bonus: bottom left you can see turbidity (cloudiness) illustrated with a black background, a home depot puck light, and a white plastic cap diffuser. The scattering from the bacteria in the water is very evident in the samples that grew vs the killed control. Bottom right is the scattering quantified with a digital photo app.
One thing we might notice here is that the amount of O2 consumption and ammonia production seem faster in the native aquarium bacteria than in the bottle bacteria product. Additionally, the ammonia production (a marker for breakdown of compounds with excess N) reaches far higher with native aquarium bacteria. Worth thinking about, but let's leave these here for now and see if they show up again later.

Ok, a 1% dilution of a bacterial growth media can activate bottle bacteria, but can we achieve that with something more plausible for an aquaria? We failed with fish food, but maybe it just wasn't rich enough? And lets look at a bunch of bottle bacteria products.

So here's Frozen Cube (Emerald Entree) 50mg/L of protein.
Treatments are:
Heat-killed: Pristine, Waste Away, MicroBacter7 listed as "Pri (Ster)" etc
Raw recommended dose: WA, MB7, Aq water, my old (maybe dead) bottle of WA, Pri
Two additional controls got nothing.
I ground up (mortar and pestle) a measured amount (50mg/L protein) to a paste and added to Instant Ocean. Agitated the bottle to make it uniform as I split into all the ~20mL tubes. Heat-killed with 30 min covered boil, and added bacteria treatments.

Ooh boy! we asked for activity, and here we've got it by the buckets. Every duplicate of every product came alive while the killed versions of the same products act just like blank controls. Even my old bottle of WA, that I was worried about, still perked up. All consumed O2, and all did....stuff to the ammonia reading. Digestion of Nitrogen-rich compounds could release excess N, and building biomass could consume N, so both are possible.
Total ammonia test chemistry reacts with ammonia and proteins and aminos, so the initial levels / controls are not really much ammonia - but interfering N in proteins etc. The increase of ammonia after digestion is real and almost all actually ammonia.

Notice that again the native aquarium bacteria were the most aggressive oxidizers of carbon - O2 in sealed tubes was fully depleted - and the largest producers of ammonia. (The 2nd WA sample looks nothing like any other bacterial product sample and looks a lot like Aq bacteria so lets think of it as likely contamination - oops.) Other than that, it's quite interesting how different the bottle product results look from the native Aq bacteria when presented with the same nutrients.

So bottled bacteria products absolutely contain viable bacteria that can grow. But so far we've seen growth on rich laboratory bacterial media and a fish food level of ~50mg/L protein - when I only feed an average of ~3mg/L protein to my tank on a daily basis. (And earlier 30mg/L of flake - 16mg/L protein showed no activity.)

To my mind these are the big Qs after what we've seen so far.

• Is there a "richness gap" between what nutrients seem needed to activate the bacteria and what exists in our tanks?
• Does native Aquarium bacteria more thoroughly decompose food matter than bottle bacteria?
• Is there a multi-day lag time of inactivity with bottle bacteria compared to native bacteria?

Next: We'll see if some of those questions are within our reach and take a look at specific bottle bacteria claims to try to match those to experimental situations.

Great conclusion! Those darn big three questions got me thinking.

1) In these experiments, the observed metabolism of a consortium of microorganisms is being compared to spores or a limited number of species. That is neither good nor bad, just a clarification. I do wonder if the consortium make up changes over the course of the experiment. @AquaBiomics would be a good resource to tap into if Eli is interested in helping.

2) Spore germination and the growth lag period as bacteria gear up to assimilate a new food, puts the bottled bacteria at a disadvantage in a five day race against microorganisms from the aquarium that are “up and running“. This observation might be a ”so what” observation because that is exactly what happens in the aquarium.

3) Is it known whether these bottled bacteria do well growing in a biofilm rather than in the water column? And should it really matter?

4) These bottled bacteria products make some exciting claims. I was therefore surprised that they don’t germinate in the presence of fish food and required essentially dilute “beef broth” (LB medium) to germinate. I wonder what they eat when they wake up when there is any beef broth around?

5) When comparing the metabolic capabilities of bottled bacteria to the aquarium consortium, are the bottled bacteria going to be cultured first, letting them get a running start?

 

[taricha]

Also why these products are pointless. A bottle of sleeping bacteria isn't going to alter the balance in your tank.

Oh and adding bacteria to an already establish aquarium is a gimmick.

While I try to dissuade new reefers from wasting their money on multiple infusions of 'Bottle Bac'

Come on, y'all. Have you read the bottles? Powerful stuff!

In seriousness, for some products the positive observations are high enough that it exceeds what I think the usual placebo power is in this hobby. If the effects are real, it ought to be measurable.

Now not sure what the end goal of your experiment is

mostly to understand what aquarium bacteria do in great detail, so the interventions we make to target bacteria can be something other than random.

 

[Alenya]

Conclusion of Part 1: Waking up Sleeping Beauty (for real this time)

When we left off in post 1 we had tried fish flake and skimmate, and our bottled bacteria products did precisely nothing. Not consuming PO4 to build biomass, not consuming Oxygen (oxidizing Carbon for energy), and not breaking down proteins etc to release N, seemingly not waking up.
Makes us worried if there are viable bacteria in the bottles. Let's go way out of the box and try something that isn't an aquarium thing at all, but a common bacterial growth medium - LB media.

Tank water was spiked to 1% LB media, heat-killed and added treatments of killed Waste Away "WA(st)", raw Waste Away "WA", or Aquarium water "Aq"
150mL flasks (day 3 measurements in extra 20mL test tubes). "heat-killed" media and Waste Away were done by covered 30 min boiling in tightly covered pot. In order to allow more growth with less restriction from O2 in this rich media, 10% air space was left in each sample, so O2 could diffuse in slowly.

And Voila! we have vigorous activity. By day 5 we have complete O2 consumption by the bottled (and aquarium) bacteria and plenty of Ammonia production. Bonus: bottom left you can see turbidity (cloudiness) illustrated with a black background, a home depot puck light, and a white plastic cap diffuser. The scattering from the bacteria in the water is very evident in the samples that grew vs the killed control. Bottom right is the scattering quantified with a digital photo app.
One thing we might notice here is that the amount of O2 consumption and ammonia production seem faster in the native aquarium bacteria than in the bottle bacteria product. Additionally, the ammonia production (a marker for breakdown of compounds with excess N) reaches far higher with native aquarium bacteria. Worth thinking about, but let's leave these here for now and see if they show up again later.

Ok, a 1% dilution of a bacterial growth media can activate bottle bacteria, but can we achieve that with something more plausible for an aquaria? We failed with fish food, but maybe it just wasn't rich enough? And lets look at a bunch of bottle bacteria products.

So here's Frozen Cube (Emerald Entree) 50mg/L of protein.
Treatments are:
Heat-killed: Pristine, Waste Away, MicroBacter7 listed as "Pri (Ster)" etc
Raw recommended dose: WA, MB7, Aq water, my old (maybe dead) bottle of WA, Pri
Two additional controls got nothing.
I ground up (mortar and pestle) a measured amount (50mg/L protein) to a paste and added to Instant Ocean. Agitated the bottle to make it uniform as I split into all the ~20mL tubes. Heat-killed with 30 min covered boil, and added bacteria treatments.

Ooh boy! we asked for activity, and here we've got it by the buckets. Every duplicate of every product came alive while the killed versions of the same products act just like blank controls. Even my old bottle of WA, that I was worried about, still perked up. All consumed O2, and all did....stuff to the ammonia reading. Digestion of Nitrogen-rich compounds could release excess N, and building biomass could consume N, so both are possible.
Total ammonia test chemistry reacts with ammonia and proteins and aminos, so the initial levels / controls are not really much ammonia - but interfering N in proteins etc. The increase of ammonia after digestion is real and almost all actually ammonia.

Notice that again the native aquarium bacteria were the most aggressive oxidizers of carbon - O2 in sealed tubes was fully depleted - and the largest producers of ammonia. (The 2nd WA sample looks nothing like any other bacterial product sample and looks a lot like Aq bacteria so lets think of it as likely contamination - oops.) Other than that, it's quite interesting how different the bottle product results look from the native Aq bacteria when presented with the same nutrients.

So bottled bacteria products absolutely contain viable bacteria that can grow. But so far we've seen growth on rich laboratory bacterial media and a fish food level of ~50mg/L protein - when I only feed an average of ~3mg/L protein to my tank on a daily basis. (And earlier 30mg/L of flake - 16mg/L protein showed no activity.)

To my mind these are the big Qs after what we've seen so far.

• Is there a "richness gap" between what nutrients seem needed to activate the bacteria and what exists in our tanks?
• Does native Aquarium bacteria more thoroughly decompose food matter than bottle bacteria?
• Is there a multi-day lag time of inactivity with bottle bacteria compared to native bacteria?

Next: We'll see if some of those questions are within our reach and take a look at specific bottle bacteria claims to try to match those to experimental situations.

Seeing actual scientific method, quantifiable results and process makes me very happy.

Thank you for taking the time to test and to post your findings!

 

[flampton]

Come on, y'all. Have you read the bottles? Powerful stuff!

In seriousness, for some products the positive observations are high enough that it exceeds what I think the usual placebo power is in this hobby. If the effects are real, it ought to be measurable.


mostly to understand what aquarium bacteria do in great detail, so the interventions we make to target bacteria can be something other than random.

What I am sure off is that it is the placebo effect. You buy something you want it to work. Then you pay more attention and your husbandry increases. It works!

I'll tell you that the mechanism of these products to work is that the bacterial population has to expand. After it expands it is in your aquarium consuming nutrients. The population will only crash with a change, e.g. carbon type etc... Now if it doesn't expand, well it will be pulled out. The dosages of these products will never add enough bacteria to be useful without expanding.

Now there is a caveat to the above as some of these products have other additives that could potentially be doing something observable

 

[flampton]

Okay I thought about it a little more and that one other way these might be useful is if you do change the carbon dosing method. So going from vodka to vinegar etc. But it would only need a few infusions before new populations would be established.

Now when we talk about solid carbon dosing, PHA, I.e. biopellets, What we find is that it's not very effective for the first few weeks. This is because of the slow digestion and limited strains that can utilize this substrate. For this reason I purchased two little fishies bacteria for establishing a good population of PHA utilizing bacteria. After my aquarium is up and running for a significant amount of time I'll check to see if the two little fishies product matches what is growing on my pelllets. If not that's another product idea...

 

[taricha]

Great conclusion! Those darn big three questions got me thinking.

1) In these experiments, the observed metabolism of a consortium of microorganisms is being compared to spores or a limited number of species. That is neither good nor bad, just a clarification. I do wonder if the consortium make up changes over the course of the experiment. @AquaBiomics would be a good resource to tap into if Eli is interested in helping.

2) Spore germination and the growth lag period as bacteria gear up to assimilate a new food, puts the bottled bacteria at a disadvantage in a five day race against microorganisms from the aquarium that are “up and running“. This observation might be a ”so what” observation because that is exactly what happens in the aquarium.

3) Is it known whether these bottled bacteria do well growing in a biofilm rather than in the water column? And should it really matter?

4) These bottled bacteria products make some exciting claims. I was therefore surprised that they don’t germinate in the presence of fish food and required essentially dilute “beef broth” (LB medium) to germinate. I wonder what they eat when they wake up when there is any beef broth around?

5) When comparing the metabolic capabilities of bottled bacteria to the aquarium consortium, are the bottled bacteria going to be cultured first, letting them get a running start?

1) and 2) do a good job to give some context for why the bottled bacteria might not be as aggressive on our measurable quantities as native Aq bacteria are.

5) (pre-culturing the bottle stuff) would do a good job of removing the lag time issue mentioned in 2). It wouldn't be a test of how the product is actually used - neither is sterilizing everything - but in the same way it might make a lot clearer what the bottle bac could do in theory.
There are a few people on these forums who take their bottle products, and add them to a cup of water and feed vodka / vinegar in the hopes that the product will activate quickly and multiply and hit the tank running.
So at some point I ought to do the same.
Thinking through a test....
Culture up bottle bac on LB media. Test cultured bottle stuff and Aquarium bac on food source. After X days, measure and cross-inoculate - take some of the bottle bac and add to the Aq treated sample, and take some of the Aq sample and add to the bottle bac sample.
See if either leaves behind goodies that the other can use - or if they both use up the food just in different ways.

3) (films vs water) my food tests have settled fluffy solids and the spores presumably come to rest in the food stuffs. I'd think if they wanted to form a surface colony on the food directly, then they could digest it very well that way?

1) absolutely the consortium of Aq bac changes. If you feed a bottle twice, the response the second time around (to the same food) is always notably faster.

 

[Variant]

Could the lag be cause by bacterial count versus metabolism? Perhaps some pf these large bottles of bac Hve lower concentrations?

 

[taricha]

Could the lag be cause by bacterial count versus metabolism? Perhaps some pf these large bottles of bac Have lower concentrations?

When I take these products and centrifuge, they leave a nice pellet of solid spores at the bottom of the tube. Pour off the bottle media replace with distilled water and shake to resuspend the pellet and you get a nice cloudy liquid suspension of bacteria spores.
Compare that to the aquarium water which when added is optically clear. So the number of bacteria spores that go in should certainly compare favorably to the live aquarium bacteria. (With the caveat that spores scatter light with about ~2x the optical density of germinated growing bacteria - but still, it's way more than 2x as cloudy as Aq water).
But how many of those scattering spore particles are viable, and how many wake up in a reasonable time to grow and make a dent in our measurements? Maybe it's a tiny fraction of the total and much lower than what gets in with a small volume of aquarium water?

 

[taricha]

Part 2: The Sleeper has Awakened: What does it do when it wakes?

In Part 1 we saw that just because an aquarium food source contains nutrients and native aquarium bacteria consume it, doesn't mean it'll trigger growth of bottled products (post 1), but the bottled bacteria are viable and can be awakened by some sources, even aquarium sources - very rich ones at least (post 13).
The data thus far pushes 3 big questions to the front for me.

1. Is there a "richness gap" between what nutrients seem needed to activate the bacteria and what exists in our tanks?
2. Does native Aquarium bacteria more thoroughly decompose food matter than bottle bacteria?
3. Is there a multi-day lag time of inactivity with bottle bacteria compared to native bacteria?

number 2 is the hardest, so we'll look at the others for now.

Let's check the timing response of bottle bacteria and native aquarium bacteria.
This is with LB media - the bacterial growth medium used Figure 3 but diluted to 0.5% this time instead of 1%.
Details: 20mL tubes with Aquarium water spiked to 0.5% LB Media. Heat-killed by covered pot boiling 30 minutes. Then inoculated with bacteria, sealed and opened at test time. Each data point is a replicate tube - not one sample opened on multiple days.

Figure 5: LB Media diluted to 0.5% Inoculated with Waste Away, MicroBacter7, and Aquarium Water

Oxygen consumption (from oxidized Carbon), Ammonia production (from breaking down proteins), and pH drop (CO2 and/or other acid production) all tell the same story. Every sample inoculated with aquarium water showed activity between day 0.7 and 1.7, while the Waste Away samples showed activity by day 3.7. MicroBacter7 really wasn't interested in the stuff and showed no activity even after nearly 6 days.
(PO4 is complicated because the medium likely contains a lot of P in forms other than PO4 that don't show up on a test, so decrease of PO4 could happen by bacterial growth, and increase of PO4 could happen by breakdown of the organic P forms to inorganic PO4.)

So it looks like right at a 3 day lag for the bottled bacteria compared to aquarium bacteria, which themselves had maybe a half-day lag - though that is understandable as I don't normally feed my tank laboratory bacterial growth media

Here's another test run done with a high level of fish flake (the same flake that got no response in the first post)
Details: 150mL flasks with Aquarium water and 100mg/L fish flake (53mg/L protein). Heat-killed by pressure canner. Then inoculated with bacteria, and sealed until samples withdrawn and tested at days 2, 4 & 7. This method lost sterilization after 8 days and was improved on later. I'd take it with a grain of salt, but it is consistent with other results.

Figure 6: Fish Flake 100mg/L (53mg/L protein) Inoculated with Waste Away, Pristine, Live Rock Enhance, Heat-Killed Pristine, and Aquarium Water

Oxygen consumption shows activity from Pristine and Live Rock Enhance between day 2 and 4, and from Waste Away after day 4. Ammonia production shows the exact same story - with the additional information that the aquarium bacteria releases far more ammonia from protein breakdown than the bottle products.
All this data is also consistent with a near-immediate response from aquarium bacteria but a 2-4 day lag time from the bottle bacteria products. (A separate test with same food & concentration also had MicroBacter7 showing activity at day 4.)

This gives a different interpretation to the lack of activity in the earlier fish flake test in post 1, Fig 1
Maybe the issue is a long delay and not just a lack of richness. Even if the fish flake (16mg/L protein) in that first test were rich enough, the hold time for that first test was only 2.5 days.

So where do we stand?
Thus far, in every sample that wakes the bottled bacteria (Fig 3, 5 & 6), the activity is only detectable with a 2-4 day lag behind the native aquarium bacteria. And that's with bacterial growth media or 3 weeks' worth of fish food (50mg/L protein).
On the "richness" question, a mere 5 days' worth of fish food (16mg/L protein) showed no activity in bottle bacteria in 2.5 days, but it's unknown if it wasn't enough food, or if hold time was too short and activity would've appeared after 3-4 days.

 

[terraincognita]

One note I'll make really quick I haven't been able to read every Reply But I think a major component that is missing and isn't quite the same is the fact that the bacteria need something to host in and I do not think you are performing any of these test and providing something for the bacteria to host in the water if I'm not mistaken?

I.e. you can't have a Saltwater aquarium without some type of rock, bio-filter media.

I'm not sure if I'm missing out on some major point here.

But from what I'm understanding these tests really are not showing anything without providing a proper environment?

Like technically you're not really activating any of the bacteria, so it's just like putting it from one bottle into another bottle? and then adding fish food? I'm not trying to hate on this, I just would like to see test done with some bio media or something that could be sterile when added. but I think important for reef keeping sakes.

Otherwise these tests have nothing to do with the keeping of fish or a reef.

 

[Dan_P]

One note I'll make really quick I haven't been able to read every Reply But I think a major component that is missing and isn't quite the same is the fact that the bacteria need something to host in and I do not think you are performing any of these test and providing something for the bacteria to host in the water if I'm not mistaken?

I.e. you can't have a Saltwater aquarium without some type of rock, bio-filter media.

I'm not sure if I'm missing out on some major point here.

But from what I'm understanding these tests really are not showing anything without providing a proper environment?

Like technically you're not really activating any of the bacteria, so it's just like putting it from one bottle into another bottle? and then adding fish food? I'm not trying to hate on this, I just would like to see test done with some bio media or something that could be sterile when added. but I think important for reef keeping sakes.

Otherwise these tests have nothing to do with the keeping of fish or a reef.

Are you suggesting that bottled bacteria will not grow on plastic or glass surfaces? That if there was a ”natural” surface the bacteria would settle, form a biofilm and grow?

If so, I wonder about the same thing. Are bottled bacteria a fussy benthic type