Help with experiment parameters discussion on DOC saltwater demand and microbial evaluation.

[MnFish1]

Tw aquabiomics may want to partner with you an the experiment I would suggest this

 

[sixty_reefer]

There is simply no way to make conclusions on this post except 10 hypotheticals which will not help imho. I applaud your method. And though I don’t necessarily think Dan p s experiments can adequately explain in a full tank with fish etc. I would consider going along his route with smaller volumes and more experimental tanks. Don’t take this as a criticism. It’s not meant that way. But I think it’s the only way to do what you’re trying to do. Note this is also not a criticism against Dan p. I just wonder how well these micro aquaria translate to mature 500 gallon tanks

I have lots of respect for you to ever take your criticism the wrong way mate.
What’s happening in here is not much different from biofloc water treatment. There is many articles on the concept although I wasn’t aware that it was possible to do it in a controlled manner in a marine environment without the cloudy water. The outcome seems to be much similar.
I’ve done it in the past using different forms of carbohydrates with a great challenge in keeping conditions suitable for a reef aquarium.
There isn’t any method out there that allows a clean culture and harvesting (protein skimmer cup) of bacteria and single cell marine zooplankton (smallest zooplankton in saltwater). I’m happy with the outcome so far as potential I could use this to feed hard to keep NPS and crinoids and obviously any others photosynthetic small polyp coral.
My theory was based on mimicking seawater conditions in certain environments that would allow the basic bottom chains of feeding to develop.

I was going to expand the test into removing some nutrients and see the effects but as even if the outcome was to be as expected, it wouldn’t change much in the hobby.

 

[sixty_reefer]

Tw aquabiomics may want to partner with you an the experiment I would suggest this

Hi have thought of doing a test at some point just to see how different the test could be from a normal system.

 

[sixty_reefer]

Hi - you have still not answered my questions or Dan's. In any case, You appear to be counting Protozoa and hope that parameters stay the same - Forgive, me but what is the correlation to explaining how carbon dosing affects phosphate levels?

Just a observations, carbon dosing usually thrives some phosphate down.

Repeating prior thoughts and questions in case you missed
Thoughts:

-Biofilm accounts for 90-95% of the biomass (excluding fish and coral if they are present).

Yes although my talks always mention exploring the pelagic biofilm.

-No quantification of C
-You can't measure DOC

I’ve measure it it’s in the OP

-makeup of biofilm and water column is significantly different.

Agreed

-The skimmer samples are not representative of the water column. They have a different concentration and likely show different bioactivity.

They a indicate of how much I can harvest from a small tank

If ~5% of the biomass is in the water column and it greatly differs from the bulk of the biomass on surfaces, I am not sure what is being quantified to what conclusion.

There is a test that can be used in a later stage to quantify the biomass in the water column

Is there a hypothesis?
What are the controls?
Can you explain how the results intend to be interpreted?

Every one has their own mind, I can only post results

Dan_P (among other things asked):
For example were you expecting the number of protozoa to decline or the species to change?

I would hope that I could increase and decrease the cultures by just increasing or decreasing the available C and N

 

[BeanAnimal]

Just a observations, carbon dosing usually thrives some phosphate down.

This doesn't explain how carbon dosing in your setup is relevant to phosphate levels or Protozoa. Could you clarify the relationship you're proposing?

Yes although my talks always mention exploring the pelagic biofilm.

That is not an answer to the question and doesn't clarify the relevance of focusing on the pelagic bacteria when the majority of microbial biomass resides in surface biofilms.

"Your talks?" -I am also not sure what your meaning is here. Are you an authoritative expert that can be cited on this matter or did you mean something else?

I’ve measure it it’s in the OP

You mentioned a home-derived source for the carbon, but how are you quantifying DOC without the proper tools? Please clarify.

Agreed

It was not a question. The biology in the biofilm, water column and skimmate are different and the biofilm is 90-95% of the biomass in the system. You are measuring what is in the skimmate. How does this correlate to the vast majority of the biology in the tank that will be consuming nutrients?

Moreover, the skimmer is not only selectively removing a subset of what is biologically presented to it, but everything from barometric pressure to water chemistry to the biofilm on the skimmer surfaces affect (greatly) its performance and what it pulls out in what quantity.

They a indicate of how much I can harvest from a small tank

So the experiment is to determine how to most efficiently dose the aquarium to produce Protozoa in skimmate?
This sounds more like a harvesting experiment than a biological observation. Can you clarify?

There is a test that can be used in a later stage to quantify the biomass in the water column

Citation please supporting this claim please.

Every one has their own mind, I can only post results

That is not an answer to the question. I don't see where you’ve clearly defined the experiment parameters or controls. Providing that framework would help everyone understand your process better.

I would hope that I could increase and decrease the cultures by just increasing or decreasing the available C and N

That's an assumption, but without defining how you plan to measure and interpret those increases or decreases, it remains speculative (at best).

 

[sixty_reefer]

This doesn't explain how carbon dosing in your setup is relevant to phosphate levels or Protozoa. Could you clarify the relationship you're proposing?


That is not an answer to the question and doesn't clarify the relevance of focusing on the pelagic bacteria when the majority of microbial biomass resides in surface biofilms.

"Your talks?" -I am also not sure what your meaning is here. Are you an authoritative expert that can be cited on this matter or did you mean something else?


You mentioned a home-derived source for the carbon, but how are you quantifying DOC without the proper tools? Please clarify.


It was not a question. The biology in the biofilm, water column and skimmate are different and the biofilm is 90-95% of the biomass in the system. You are measuring what is in the skimmate. How does this correlate to the vast majority of the biology in the tank that will be consuming nutrients?

Moreover, the skimmer is not only selectively removing a subset of what is biologically presented to it, but everything from barometric pressure to water chemistry to the biofilm on the skimmer surfaces affect (greatly) its performance and what it pulls out in what quantity.


So the experiment is to determine how to most efficiently dose the aquarium to produce Protozoa in skimmate?
This sounds more like a harvesting experiment than a biological observation. Can you clarify?


Citation please supporting this claim please.


That is not an answer to the question. I don't see where you’ve clearly defined the experiment parameters or controls. Providing that framework would help everyone understand your process better.


That's an assumption, but without defining how you plan to measure and interpret those increases or decreases, it remains speculative (at best).

And that’s why this is a test, seen all you want is a discussion

 

[BeanAnimal]

And that’s why this is a test, seen all you want is a discussion

But the discussion is wandering and makes absolutely no sense. You have answered almost every question with a deflection or only tangentially relevant responses.

You have not defined what you are testing or how you are testing or how you intend to interpret the results of your test.

Can you do that for us so that we may help you?

 

[sixty_reefer]

But the discussion is wandering and makes absolutely no sense. You have answered almost every question with a deflection or only tangentially relevant responses.

You have not defined what you are testing or how you are testing or how you intend to interpret the results of your test.

Can you do that for us so that we may help you?

At the moment you looking at the test from a nutrient control perspective if you would like to understand or help what I’m doing you will have to look at it with the perspective that I’m making a culturing system similar to a phytoplankton or rotifers culture, the goal here is to use 3 chemicals to propagate and harvest bacteria and single cell zooplankton wile keeping the parameters similar to a reef environment.

The conditions of the tank need to be similar to a reef environment due to at some point a subject test like a crinoid or a carnation coral will have to be added to the tank to evaluate feeding conditions.
Ideally a secondary tank would be most effective to keep the target feeding subject and add the food harvested from the culturing tank to it.

The goal for the first tank (culturing tank) is to mimic deep seawater nutrient demand that can be found in their natural habitat. Were carbohydrates from macro and micro algae’s will be floating in the strong currents of this particular environments and attached to the carbon there will be bacteria feeding on it and single cell zooplankton feeding on the bacteria, creating a floating biofilm that could potentially be the food for the main subject (crinoids and NPS).
In a normal reef tank a sand bed and rock could be detrimental for this floating biofilms by trapping them within the sand or rock therefore I’ve decided to make the display bare and enough flow to keep the particles in suspension.
The only biofilms intend to be evaluated in this experiment is the ones floating as this one’s the ones that could potentially work as a feed for the subject in question.

The tool that can be used to measure the culture density on the water column is called a Imhof cone, this cone gives you a general idea of the density of the floating biofilms.

The protein skimmer is doing what is designed to do and collecting this biofilm from the water column, there isn’t any fish biological load in this tank therefore all it’s collecting at this point in time is this biofilms that carry on developing in the skimmer cup.
Looking at the content of the protein skimmer cup under the microscope at this stage is just to have a indication that there is carbon, bacteria and single cell zooplankton in suspension in the water column.

There is no need for control, as this is one of those experiments that it will work or just fail, I don’t believe that there is many tanks out there encouraging the growth of floating biofilms with the carbon or the conditions used in this experiment that would need a comparison to.



Takin the above explanation, what would the critics be?

 

[Dan_P]

At the moment you looking at the test from a nutrient control perspective if you would like to understand or help what I’m doing you will have to look at it with the perspective that I’m making a culturing system similar to a phytoplankton or rotifers culture, the goal here is to use 3 chemicals to propagate and harvest bacteria and single cell zooplankton wile keeping the parameters similar to a reef environment.

The title could have been worded something like “help with development of bacteria and zooplankton culturing” instead of​

Help with experiment parameters discussion on DOC saltwater demand and microbial evaluation.​

Kinda wasted time on the wrong purpose

 

[sixty_reefer]

The title could have been worded something like “help with development of bacteria and zooplankton culturing” instead of​

Help with experiment parameters discussion on DOC saltwater demand and microbial evaluation.​

Kinda wasted time on the wrong purpose

From a hobby perspective if I have the word zooplankton most would have commented with just add phytoplankton, most of the thread would of been on the different types of zooplankton in the sea and still not get the help I needed.
what I needed to understand was how different forms of C ( more in particular carbohydrates) created life in a saltwater aquatic environmental and their nutrient demands as that’s what’s consuming nutrients in seawater. Water itself doesn’t need nutrients.
The end motive wasn’t important for the discussion from my perspective although I have to mention now for some context and get back to the focus of the experiment.

From what I’ve seen so far it appears that the biofilm is created around the carbon particles, I didn’t knew that previously to start this experiment. Possible meaning that the water column could have as much biofilm as the surface area of a aquarium depending on how much particulate carbon is in the water column.

 

[sixty_reefer]

In addition @Dan_P there is a large movement in the forum that believes that to increase bacterial population, we just need to introduce them the tank.
In my views I’ve always believed that we can’t increase those numbers without also introducing the nutrients required to get those numbers up.

I don’t think I can illustrate that though with the limited resources I have available to me, hopefully someone with more resources in future could look into that.

 

[BeanAnimal]

At the moment you looking at the test from a nutrient control perspective if you would like to understand or help what I’m doing you will have to look at it with the perspective that I’m making a culturing system similar to a phytoplankton or rotifers culture, the goal here is to use 3 chemicals to propagate and harvest bacteria and single cell zooplankton wile keeping the parameters similar to a reef environment.

I (we) are looking at it from the perspective that you presented it. You initial posts were regarding nutrient management and limitations and you shifted to single-cell zooplankton when that conversation become tough. Your intent and focus have not been clear.

Nonetheless you are still referring to this as a "test" but have dismissed the need for a protocol or controls and I am (honestly) still unclear on the actual hypothesis.

The conditions of the tank need to be similar to a reef environment due to at some point a subject test like a crinoid or a carnation coral will have to be added to the tank to evaluate feeding conditions.
Ideally a secondary tank would be most effective to keep the target feeding subject and add the food harvested from the culturing tank to it.

Those statements seem contradictory. If the coral lives in the culture, the tank must support coral. If the coral lives in a separate system, the conditions of the culturing tank don't need to mimic a reef, and they should be optimized for propagation and not coral survival.

The goal for the first tank (culturing tank) is to mimic deep seawater nutrient demand that can be found in their natural habitat. Were carbohydrates from macro and micro algae’s will be floating in the strong currents of this particular environments and attached to the carbon there will be bacteria feeding on it and single cell zooplankton feeding on the bacteria, creating a floating biofilm that could potentially be the food for the main subject (crinoids and NPS).

Your premise here is very flawed. The carbohydrates you speak of (algae derived) are generated and consumed in the shallows, not in deep water. Zooplankton may dive to avoid predation, but hey feed in shallow waters where their food source is produced.

Moreover, the nutrients at depth (in this context) are the result of decomposition (sinking particles). So the carbon sources you describe as "deep water" are actually shallow water.

Crinoids and NPS primarily use these decomposed suspended particles for food, not bacteria (or Protozoa). So your proposed floating biofilm is not the primary food source. Even if it was, I don't see any controls or test protocols that would be able to identify or quantify this mechanism. You are counting "Protozoa" in a skimmer cup.

In a normal reef tank a sand bed and rock could be detrimental for this floating biofilms by trapping them within the sand or rock therefore I’ve decided to make the display bare and enough flow to keep the particles in suspension.
The only biofilms intend to be evaluated in this experiment is the ones floating as this one’s the ones that could potentially work as a feed for the subject in question.

Ignoring any debates about biological filtration, you are assuming that the source of these floating biofilms is from the water column and not the vastly increased surface area created by the substrate. You have not provided any method or controls to confirm or reject this assumption.

Most biofilms are surface attached, not free floating in the water column. Do you have supporting evidence otherwise?

The tool that can be used to measure the culture density on the water column is called a Imhof cone, this cone gives you a general idea of the density of the floating biofilms.

But you are not evaluating the biofilm (attached or floating) or the water column in general, but rather the skimmate. To that end, the Imhoff Cone is typically used to measure suspended solids, not bacterial density.

The protein skimmer is doing what is designed to do and collecting this biofilm from the water column, there isn’t any fish biological load in this tank therefore all it’s collecting at this point in time is this biofilms that carry on developing in the skimmer cup.
Looking at the content of the protein skimmer cup under the microscope at this stage is just to have a indication that there is carbon, bacteria and single cell zooplankton in suspension in the water column.

To what end? As long as the system is not sterile you are going to find microbial acitvity. Quantifying anything beyond that point is not supported due to the numerous variables involved. Again, this "test" lacks any meaningful controls.

There is no need for control, as this is one of those experiments that it will work or just fail, I don’t believe that there is many tanks out there encouraging the growth of floating biofilms with the carbon or the conditions used in this experiment that would need a comparison to.

if there is no hypothesis, no protocol and no controls then what is "working" vs "failing"? You can't quantify results and can't compare different variables without controls.

 

[sixty_reefer]

I (we) are looking at it from the perspective that you presented it. You initial posts were regarding nutrient management and limitations and you shifted to single-cell zooplankton when that conversation become tough. Your intent and focus have not been clear.

Nonetheless you are still referring to this as a "test" but have dismissed the need for a protocol or controls and I am (honestly) still unclear on the actual hypothesis.


Those statements seem contradictory. If the coral lives in the culture, the tank must support coral. If the coral lives in a separate system, the conditions of the culturing tank don't need to mimic a reef, and they should be optimized for propagation and not coral survival.


Your premise here is very flawed. The carbohydrates you speak of (algae derived) are generated and consumed in the shallows, not in deep water. Zooplankton may dive to avoid predation, but hey feed in shallow waters where their food source is produced.

Moreover, the nutrients at depth (in this context) are the result of decomposition (sinking particles). So the carbon sources you describe as "deep water" are actually shallow water.

Crinoids and NPS primarily use these decomposed suspended particles for food, not bacteria (or Protozoa). So your proposed floating biofilm is not the primary food source. Even if it was, I don't see any controls or test protocols that would be able to identify or quantify this mechanism. You are counting "Protozoa" in a skimmer cup.


Ignoring any debates about biological filtration, you are assuming that the source of these floating biofilms is from the water column and not the vastly increased surface area created by the substrate. You have not provided any method or controls to confirm or reject this assumption.

Most biofilms are surface attached, not free floating in the water column. Do you have supporting evidence otherwise?


But you are not evaluating the biofilm (attached or floating) or the water column in general, but rather the skimmate. To that end, the Imhoff Cone is typically used to measure suspended solids, not bacterial density.


To what end? As long as the system is not sterile you are going to find microbial acitvity. Quantifying anything beyond that point is not supported due to the numerous variables involved. Again, this "test" lacks any meaningful controls.


if there is no hypothesis, no protocol and no controls then what is "working" vs "failing"? You can't quantify results and can't compare different variables without controls.

I must not be reading correctly, I thought you wanted to “help”, I’m still not to sure how your current persistent criticism is meant to “help me” in any way.

 

[BeanAnimal]

From what I’ve seen so far it appears that the biofilm is created around the carbon particles, I didn’t knew that previously to start this experiment.

Biofilms typically form where there is surface area and a nutrient source. It's fundamental to microbial biology, so it’s surprising that after (3) threads discussing bacteria, this seems new to you.

I’m unsure how your experiment actually proved this, but yes, it is basic to biofilm formation.

Possible meaning that the water column could have as much biofilm as the surface area of a aquarium depending on how much particulate carbon is in the water column.

For that to happen, the suspended carbon would need to have surface area comparable to the rest of the substrate and hard surfaces in the tank. Are you trying to create a bacterial sludge for harvesting, or a viable reef system?

I must not be reading correctly, I thought you wanted to “help”, I’m still not to sure how your current persistent criticism is meant to “help me” in any way.

Criticism here isn't meant to be negative for the sake of it.

Several of us (in three threads) have been trying to help you move forward. We can't do that without addressing the gaps in your method and understanding. However, each time we address something you either dismiss it, change direction or introduce new tangent concepts or ideas, as if we are the people who don't understand the concepts being discussed.

Basic points are being pressed because your explanations are evolving further and further from the point. These exchanges are not going in circles, but rather spiraling away from very basic and important information and toward irrelevant concepts that are at best loosely related to the point.

Unfortunately, we can't help you because you portray yourself as already having the answers and avoiding or dismissing feedback.

With that, I’ll step away from this discussion and wish you the best with your research.

 

[sixty_reefer]

T

Biofilms typically form where there is surface area and a nutrient source. It's fundamental to microbial biology, so it’s surprising that after (3) threads discussing bacteria, this seems new to you.

I’m unsure how your experiment actually proved this, but yes, it is basic to biofilm formation.


For that to happen, the suspended carbon would need to have surface area comparable to the rest of the substrate and hard surfaces in the tank. Are you trying to create a bacterial sludge for harvesting, or a viable reef system?



Criticism here isn't meant to be negative for the sake of it.

Several of us (in three threads) have been trying to help you move forward. We can't do that without addressing the gaps in your method and understanding. However, each time we address something you either dismiss it, change direction or introduce new tangent concepts or ideas, as if we are the people who don't understand the concepts being discussed.

Basic points are being pressed because your explanations are evolving further and further from the point. These exchanges are not going in circles, but rather spiraling away from very basic and important information and toward irrelevant concepts that are at best loosely related to the point.

Unfortunately, we can't help you because you portray yourself as already having the answers and avoiding or dismissing feedback.

With that, I’ll step away from this discussion and wish you the best with your research.

Thank you, I’m not sure if I will be able to finish this research without you criticising it.

 

[sixty_reefer]

Last update on this thread and i’ll reopen phase two in new thread, video showing the pelagic biofloc so far.

 

[Dan_P]

In addition @Dan_P there is a large movement in the forum that believes that to increase bacterial population, we just need to introduce them the tank.
In my views I’ve always believed that we can’t increase those numbers without also introducing the nutrients required to get those numbers up.

I don’t think I can illustrate that though with the limited resources I have available to me, hopefully someone with more resources in future could look into that.

I see things a little differently maybe. The big hobby fad is adding unknown, sometimes known, bacteria into a system with the intent to increase bacterial diversity, not necessarily to increase the population size. I have given products supporting this fad a provisional classification of “snake oil”. I say provisional because I have seen absolutely no evidence supporting the practice of this fad but I am still waiting for the evidence, at least until January, 2027.

Increasing a bacteria population by feeding it does not need to proved nor does changing the structure of a bacteria community by changing the food source or just increasing the amount. There are plenty of examples that this happens. Knowing exactly how the bacteria community structure changes still requires sophisticated studies and that is what the hobby might need to know, though our coral seem to do well with our lack of knowledge about the system’s microbiome.

 

[sixty_reefer]

I see things a little differently maybe. The big hobby fad is adding unknown, sometimes known, bacteria into a system with the intent to increase bacterial diversity, not necessarily to increase the population size. I have given products supporting this fad a provisional classification of “snake oil”. I say provisional because I have seen absolutely no evidence supporting the practice of this fad but I am still waiting for the evidence, at least until January, 2027.

Increasing a bacteria population by feeding it does not need to proved nor does changing the structure of a bacteria community by changing the food source or just increasing the amount. There are plenty of examples that this happens. Knowing exactly how the bacteria community structure changes still requires sophisticated studies and that is what the hobby might need to know, though our coral seem to do well with our lack of knowledge about the system’s microbiome.

I know what you saying, I would propose that you experiment with the product that you have in mind on your algae tanks. We all familiar with denitrifying bacteria respiring Nitrates although we not so much familiar with bacteria assimilation of Nitrogen.
If you ever have the time for a new project I would recommend you trying it and see the difference for yourself, it’s never about the diversity that brings the benefits in using this product it is in the increase of the number of bacteria that can assimilate nitrogen.
If you were to end up with one tank with no algae or barely no algae it wouldn’t surprise me vs. Control.

If you decide in doing it keep a eye for nitrate on the tank you decide to use that product as the demand will be significantly higher vs control.

 

[Dan_P]

I know what you saying, I would propose that you experiment with the product that you have in mind on your algae tanks. We all familiar with denitrifying bacteria respiring Nitrates although we not so much familiar with bacteria assimilation of Nitrogen.
If you ever have the time for a new project I would recommend you trying it and see the difference for yourself, it’s never about the diversity that brings the benefits in using this product it is in the increase of the number of bacteria that can assimilate nitrogen.
If you were to end up with one tank with no algae or barely no algae it wouldn’t surprise me vs. Control.

If you decide in doing it keep a eye for nitrate on the tank you decide to use that product as the demand will be significantly higher vs control.

Yes, I have testing bottled bacteria on my “to-do” list.