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

[sixty_reefer]

I am in the works on a exercise to evaluate nutrient demand by saltwater in a reef tank.
The idea is to observe the microbial population in a balanced environment and evaluate how some nutrient limitation could impact those environment.

For the experiment I’m using a 50 litre AIO that was seeded with 1kg of Bio-Spheres from a mature system and 50 litres of saltwater from a mature system.

For a fishfood and fish waste replacement I’m using for now the below.

0.002 grams of a carbohydrate that should be around 40-50 ppm wend dissolved in 50 litres of saltwater daily.

A solution of different forms of nitrogen were 10ml will be equivalent at 2 ppm in 50 litres of saltwater.

A phosphate solution that 1ml is equivalent to 0.02 ppm in 50 litres of saltwater.

The idea is to use the carbohydrates daily and match it with Nitrogen solution and Phosphate solution until a equilibrium is reached in the tank to identify a possible equilibrium on all 3 solutions for saltwater demand.
Once that equilibrium is reached I was considering to evaluate the diversity in the water column and biofilm using a microscope.
Possibly using a hemocytometer to evaluate bacterial and Protozoa numbers in a balanced environment.
My idea from there would be to remove one of the 3 nutrients and observe if it would be possible to see any of the populations being affected with a nutrient being limited.


Does the main aspects of the exercise make sense? And would it be more effective to use a stain on the hemocytometer as nutrients get limited to evaluate a difference in dead cells from living cells?
I’m way out of my comfort zone here as I’ve not done this before, any help would be appreciated?

Thanks.

 

[Dan_P]

I am in the works on a exercise to evaluate nutrient demand by saltwater in a reef tank.
The idea is to observe the microbial population in a balanced environment and evaluate how some nutrient limitation could impact those environment.

For the experiment I’m using a 50 litre AIO that was seeded with 1kg of Bio-Spheres from a mature system and 50 litres of saltwater from a mature system.

For a fishfood and fish waste replacement I’m using for now the below.

0.002 grams of a carbohydrate that should be around 40-50 ppm wend dissolved in 50 litres of saltwater daily.

A solution of different forms of nitrogen were 10ml will be equivalent at 2 ppm in 50 litres of saltwater.

A phosphate solution that 1ml is equivalent to 0.02 ppm in 50 litres of saltwater.

The idea is to use the carbohydrates daily and match it with Nitrogen solution and Phosphate solution until a equilibrium is reached in the tank to identify a possible equilibrium on all 3 solutions for saltwater demand.
Once that equilibrium is reached I was considering to evaluate the diversity in the water column and biofilm using a microscope.
Possibly using a hemocytometer to evaluate bacterial and Protozoa numbers in a balanced environment.
My idea from there would be to remove one of the 3 nutrients and observe if it would be possible to see any of the populations being affected with a nutrient being limited.


Does the main aspects of the exercise make sense? And would it be more effective to use a stain on the hemocytometer as nutrients get limited to evaluate a difference in dead cells from living cells?
I’m way out of my comfort zone here as I’ve not done this before, any help would be appreciated?

Thanks.

I can share what I have learned over the past 2 years doing something similar with very small aquaria. I need to get my thoughts together first.

 

[sixty_reefer]

I can share what I have learned over the past 2 years doing something similar with very small aquaria. I need to get my thoughts together first.

That would be great if you could, I did read many of your post on the algae and biofilm research in the past. I would also appreciate if you could point out any possible defect's or ways that it could be improved with the current parameters on how I’m proposing to proceed with this experiment.

 

[Randy Holmes-Farley]

What does this part mean?

The idea is to use the carbohydrates daily and match it with Nitrogen solution and Phosphate solution until a equilibrium is reached in the tank to identify a possible equilibrium on all 3 solutions for saltwater demand.

 

[rishma]

I have spent a lot of effort trying to achieve stable nitrate and phosphate in my 100L tank using carbon dosing and measured food input. Nitrate and phosphate stay in a very tight range for a week or two…then they drift down and I have to reduce carbon dosing to let it rise again. I don’t know why that happens but the pattern is pretty clear.

Just thought I’d share for your consideration

 

[Randy Holmes-Farley]

What does carbohydrate mean to you and why use a carbohydrate?

 

[sixty_reefer]

What dies carbohydrate mean to you and why use a carbohydrate?

I try to keep up with the descriptions as they are intended, in this experiment I was looking at using a carbohydrate derived from macro algae. The use of the word carbohydrate is to exclude ethanol and acetic acid.

 

[Randy Holmes-Farley]

I try to keep up with the descriptions as they are intended, in this experiment I was looking at using a carbohydrate derived from macro algae. The use of the word carbohydrate is to exclude ethanol and acetic acid.

A high molecular weight carbohydrate is likely to have many fewer organisms that can use it than is, say, acetate. That may give you a result that may not apply to people using different organics.

 

[sixty_reefer]

A high molecular weight carbohydrate is likely to have many fewer organisms that can use it than is, say, acetate. That may give you a result that may not apply to people using different organics.

Wend you say different organics, do you mean using organic matter as a carbs supply for example?
As that has been in my mind, I wouldn’t been able to observe decomposers potentially.

 

[Randy Holmes-Farley]

Wend you say different organics, do you mean using organic matter as a carbs supply for example?
As that has been in my mind, I wouldn’t been able to observe decomposers potentially.

I mean folks dosing acetate or ethanol or the material you pick may each drive different numbers and types of bacteria and other organisms.

 

[sixty_reefer]

I have spent a lot of effort trying to achieve stable nitrate and phosphate in my 100L tank using carbon dosing and measured food input. Nitrate and phosphate stay in a very tight range for a week or two…then they drift down and I have to reduce carbon dosing to let it rise again. I don’t know why that happens but the pattern is pretty clear.

Just thought I’d share for your consideration

I may be wrong but the carbon source I’m intending to use on this series of tests may not be readily available in the water column, some may need to be broken down by bacteria or Protozoa first if I’m not wrong.

 

[sixty_reefer]

I mean folks dosing acetate or ethanol or the material you pick may each drive different numbers and types of bacteria and other organisms.

That part is intentional. I believe acetate and ethanol it’s well documented, It’s the effects of the carbohydrates intended to use in here that may not be as well known. (At least not to me)
I’m curious to see the effects on Protozoa more in particular and how it may affect their population.

 

[rishma]

I may be wrong but the carbon source I’m intending to use on this series of tests may not be readily available in the water column, some may need to be broken down by bacteria or Protozoa first if I’m not wrong.

Did you intend to respond to my post or was your comment related to something else? I don’t understand how your response is at all related to the observation I shared. That’s ok, I don’t have anything further to contribute. Best of luck.

 

[sixty_reefer]

Did you intend to respond to my post or was your comment related to something else? I don’t understand how your response is at all related to the observation I shared. That’s ok, I don’t have anything further to contribute. Best of luck.

Yeah, in short I was trying to say that it’s not to difficult to balance a tank with the carbohydrates mentioned.

 

[Dan_P]

That would be great if you could, I did read many of your post on the algae and biofilm research in the past. I would also appreciate if you could point out any possible defect's or ways that it could be improved with the current parameters on how I’m proposing to proceed with this experiment.

Working on a response now. i need help with some definitions?

1) The idea is to use the carbohydrates daily and match it with Nitrogen solution and Phosphate solution until a equilibrium is reached in the tank

What does equilibrium refer to?

2) The idea is to observe the microbial population in a balanced environment and evaluate how some nutrient limitation could impact those environment.

What does a balanced environment refer to?

3) Once that equilibrium is reached I was considering to evaluate the diversity in the water column and biofilm

There are many biofilms in an aquarium. Was there one in particular being observed?

Thanks!

 

[sixty_reefer]

Working on a response now. i need help with some definitions?

1) The idea is to use the carbohydrates daily and match it with Nitrogen solution and Phosphate solution until a equilibrium is reached in the tank

What does equilibrium refer to?

here, I mean after adding all 3 different nutrients, get to a point we’re nitrate and phosphate stays constant without raising or decreasing my residual nitrate and phosphate. Some refer it as maturity others as balance although I believe the meaning is similar.

2) The idea is to observe the microbial population in a balanced environment and evaluate how some nutrient limitation could impact those environment.

What does a balanced environment refer to?

My idea here is to place 3 or 4 microscopic slides around the tank now and once the tank reaches the above conditions evaluate the biofilm developed on it.
I would like to possibly be able to evaluate if some bacteria or Protozoa is lost if we were to bottom out phosphate for example, this should hopefully allow to differentiate if there is any visible microbial changes from a system it’s balanced to a system that has a nutrient depletion.

I’ve been considering using this technique below to identify possible amounts of certain organisms and to see if I would observe more dead cells in a environment that is phosphate depleted vs a environment that has balanced nutrients for example.



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3) Once that equilibrium is reached I was considering to evaluate the diversity in the water column and biofilm

There are many biofilms in an aquarium. Was there one in particular being observed?

Thanks!

I’ve never looked into one under the microscope, still not to sure on what I’m going to encounter. Although I was going to try and focus on easy to identify Protozoa and bacteria for a start.

I think I would concentrate in a particular organism more if I was to notice more impact on them in a phosphate or nitrate depleted environment.

 

[Dan_P]

That would be great if you could, I did read many of your post on the algae and biofilm research in the past. I would also appreciate if you could point out any possible defect's or ways that it could be improved with the current parameters on how I’m proposing to proceed with this experiment.

Forgot to ask: will the aquarium be illuminated?

 

[sixty_reefer]

Forgot to ask: will the aquarium be illuminated?

No, I’ve been doing it dark for now to reduce photosynthetic interference, I also don’t have any aragonite in the system to remove their absorption of phosphate that could interfere with results.

The basic concept in the system comes from most systems being carbon balanced at inorganic and organic level.

 

[Dan_P]

That would be great if you could, I did read many of your post on the algae and biofilm research in the past. I would also appreciate if you could point out any possible defect's or ways that it could be improved with the current parameters on how I’m proposing to proceed with this experiment.

Here are some initial thoughts. I have a few more ideas but lets see if how I am thinking about the experiment is useful.

The model system,1 kg of used biospheres in 50 liters of aquarium water in an unlit aquarium, contains a microorganism community that developed under conditions that were quite different from those used in the experiment. Obvious community disruptions will be the bacteria-algae and protozoa-algae interactions. I am thinking this will have a substantial effect on the microorganism community structure (species and number) because organisms will die or change their diet and compete with other organisms. The microorganism community will also change because their new diet will not be equally useful to all organisms. Some will adapt, others will diminish in numbers. The amount of food available to the microorganism community during the experiment start up might not be sufficient to sustain the initial bio load. This is one aspect of the carrying capacity of the experimental system that might be exceeded and cause a substantial shift in the microorganism community structure. Other examples of where the carrying capacity can be exceeded are oxygen availability and waste removal rate (e.g., accumulation of dissolved organic matter). As the experimental system develops, the bio load will approach the carrying capacity in other parameters. This could cause another major community structure change. I am thinking that we might have a difficult time attributing a link between an experiment treatment and microorganism community structure.

The next idea concerns observing microorganism communities.

 

[sixty_reefer]

Here are some initial thoughts. I have a few more ideas but lets see if how I am thinking about the experiment is useful.

The model system,1 kg of used biospheres in 50 liters of aquarium water in an unlit aquarium, contains a microorganism community that developed under conditions that were quite different from those used in the experiment. Obvious community disruptions will be the bacteria-algae and protozoa-algae interactions. I am thinking this will have a substantial effect on the microorganism community structure (species and number) because organisms will die or change their diet and compete with other organisms. The microorganism community will also change because their new diet will not be equally useful to all organisms. Some will adapt, others will diminish in numbers. The amount of food available to the microorganism community during the experiment start up might not be sufficient to sustain the initial bio load. This is one aspect of the carrying capacity of the experimental system that might be exceeded and cause a substantial shift in the microorganism community structure. Other examples of where the carrying capacity can be exceeded are oxygen availability and waste removal rate (e.g., accumulation of dissolved organic matter). As the experimental system develops, the bio load will approach the carrying capacity in other parameters. This could cause another major community structure change. I am thinking that we might have a difficult time attributing a link between an experiment treatment and microorganism community structure.

The next idea concerns observing microorganism communities.

I totally see your points and I really appreciate you taking the time.

Unfortunately the experiment is not going the way most folks would think it would.

Taking that I’ve started this system around two weeks ago it is already showing signs of stability.

On the 10 of the October my parameters were as follow.

No3 5 mg/l
Po4 0.4 mg/l
Kh 8
Ph 8
O2 10 mg/

At that date my daily dose was

0.002 grams DOC
4 ml of nitrogen solution
0 ml of phosphates

Since then, dosage has been consistent.

Today the on the 16 of October

No3 5 mg/l
Po4 0.4 mg/l
Kh 9.5
Ph 8.2
O2 10 mg/l

At first observation, phosphate has been stable at 0.4 mg/l without any additions since the 10 of October. (Barely any phosphate has been used in the last 6 days) there was a exception on week one most likely due to colonisation of the surface area.

The daily consumption of nitrogen/nitrogen has been around 0.8 mg/l

There has not been any impact on oxygen the colour has darkened today so probably above 10 mg/l

Under the microscope, on the protein skimmer content, I can see that there is many different types of bacteria and Protozoa flourishing in the tank that leaves space for discussion if diversity is really better, so far I can’t understand the ph rise in 0.2 and the Kh rise in 1.5 as I don’t have any photosynthetic organisms in the system (no water changes have been performed). In addition the DOC used has had no impact on phosphate since the system has been stable or 6 days.