When and why did dosing nitrates start?

[Dan_P]

I have never said that - I have only said that if they get lack easy accessible food for low energy cost - they are able to change food supplier. Let us put it this way - my nearest shop where I can get potatoes is 500 m downhill from me. It cost me rather little energy (read work) to bike down there and buy my potatoes - the energy cost comes when I should uphill again . But if the potatoes is out of stock in that shop - I have to go around 9000 m to the next shop (back and forward) and get the dam uphill in the end too in order to get my potatoes. If both shops are filled up with potatoes - which should you chose - I know my answers. But I can do it more effective if I need to go to the other shop - I can take my flying mat (read KIA Picanto) and drive there - it will be more effective but cost more energy. So - then my neighbors see me open the garage - they know - there is no more potatoes in the nearest shop

Sincerely Lasse

This is the line that I question

“But it cost energy and it is easier to get P and N in the water column but if it is empty there - they form mats and get their nutrients from sediments (even inorganic) and from organic matter”

My point is that I have not read that cyanobacteria form mats to go looking for nutrients, the reason being is they do not have sufficient nutrients to generate the biomass to create those ugly mats we all hate. Food first, then mat formation.

 

[Lasse]

My point is that I have not read that cyanobacteria form mats to go looking for nutrients, the reason being is they do not have sufficient nutrients to generate the biomass to create those ugly mats we all hate. Food first, then mat formation.

Many organisms user their reserves when they go into nutrient starvation and change strategy. Well Known are many plants that normally use vegetative reproduction (cost very little energy) that start a very costly sexual reproduction strategy (cost a lot of energy) when they run into nutrient deficiency.

It could easily be the same when cyanobacteria get into starvation - and in that case - it is much easier because - IMO - they only have to redirect the carbohydrate production from the photosynthesis to be external instead of internal. And they need no P and N in order to let the photosynthesis create mats of mostly hydrocarbons.

Sincerely Lasse

 

[Lasse]

There is species of cyanobacteria that is pelagic in freshwater that if PO4 is available in the water column will use that - if not - they migrate down to the bottom during night time and grab PO4 that is released from the sediment in anaerobic and hydrogen sulphide producing areas (cost energy). Here is the link between anaerobic sediment. hydrogen sulphide production, nitrate in the water column and the occurrence of cyanobacteria blooms established. There is a huge report of this according swedish and danish lakes - but unfortunately it is only available in Swedish.

There is bacteria - as you surely know that can change between using oxygen and NO3 as electron acceptors if the favorite pathway (oxygen) not is available. And lately there have been bacteria discovered that can change between both light and chemical autotrophy, heterotrophy using all known possible electron acceptors depending on environment ( nonsulphur photosynthetic purple bacteria or effective microorganisms/bokashi)

For me - it is not surprising if an organism chose an initial more energy costly method if the question of surviving is on the table.

Sincerely Lasse

 

[Dan_P]

Many organisms user their reserves when they go into nutrient starvation and change strategy. Well Known are many plants that normally use vegetative reproduction (cost very little energy) that start a very costly sexual reproduction strategy (cost a lot of energy) when they run into nutrient deficiency.

It could easily be the same when cyanobacteria get into starvation - and in that case - it is much easier because - IMO - they only have to redirect the carbohydrate production from the photosynthesis to be external instead of internal. And they need no P and N in order to let the photosynthesis create mats of mostly hydrocarbons.

Sincerely Lasse

In general, microorganisms disperse when growing conditions become unfavorable. Cyanobacteria species that cause problems in our aquarium are very good as dispersing. This is easily demonstrated by moving a piece of cyanobacteria mat from its nutrient rich location on the sand to a petri dish with aquarium water. In my growth experiments, the tendency of the inoculum is always to disperse, not to form a mat.

As for not needing N and P to form mats, that is not quite correct. Cyanobacteria can store nitrogen and phosphorous and coupled with photosynthesis allows the cells to divide for awhile before depleting the reserves. Yes, growth continues for a short while, but the overriding tendency is dispersal because of the nutrient deficiency. Mats form because of vigorous growth and accumulation of ever lengthening filaments and occurs not under nutrient poor conditions.

 

[Sallstrom]

In general, microorganisms disperse when growing conditions become unfavorable. Cyanobacteria species that cause problems in our aquarium are very good as dispersing. This is easily demonstrated by moving a piece of cyanobacteria mat from its nutrient rich location on the sand to a petri dish with aquarium water. In my growth experiments, the tendency of the inoculum is always to disperse, not to form a mat.

As for not needing N and P to form mats, that is not quite correct. Cyanobacteria can store nitrogen and phosphorous and coupled with photosynthesis allows the cells to divide for awhile before depleting the reserves. Yes, growth continues for a short while, but the overriding tendency is dispersal because of the nutrient deficiency. Mats form because of vigorous growth and accumulation of ever lengthening filaments and occurs not under nutrient poor conditions.

Do you have an explanation on why cyanobacteria mats decrease and evetually goes away when adding nitrate to a low nutrient aquarium?

 

[taricha]

Or about ~ 29 ATP per molecule of L-glutamine. Note this is under 'perfect' conditions and in reality the yield of ATP is always lower than this.

So note step 1 you see your ammonia release and generation of some reducing equivalents

In step 2 you have your CO2 production and generation of more reducing equivalents and a little energy in the GTP

Finally the etc/oxphos you're seeing the majority of your energy production and the final reduction of O2 to H2O

Thank you for wading into the weeds on this. genuinely appreciated. Fortunate that the stoichometry here...
step 2: 10 CO2 produced
step 3: 10.5 O2 consumed
is almost the same as my wrong idea of O2 + C -> CO2. It's close enough, I'd never be able to measure the stoichometry mismatch.

One more deep-in-the-weeds point. The equations show a lot of PO4 going into the energy production of ADP to ATP. The PO4 of course be re-released when the ATP gets used.

Does the shuffling of ATP and ADP and vice versa import and export enough PO4 to be notable in the water concentration? Meaning - if I feed some straight carbon source, and the bacteria store up energy as ATP and max out their energy stores - would I see PO4 drop in the water even without the bacteria generating much new biomass?
My guess is no, that the possible pool of internal ATP is small and the cell has other ways to store larger amounts of energy. So I couldn't measure a notable PO4 difference in a water sample between "tired" bacteria and "high energy" bacteria. I wonder if that's true.

 

[taricha]

So big picture. Have low nitrates but high phosphate? L-glutamine might make a perfect carbon dosing vehicle.

Big caveat: single experiment in a stationary culture. But we know it will work to some extent but whether there is a true 50% split?

I'm a bit curious on your thoughs on what type of N source that would be most suitable in different scenarios ( like low N & P, low N & high P, cyano outbreaks, dinos, for "feeding" the coral holobiont etc).

You apparently have not read the many, many posts of folks who had no detectable nitrate, dosed it, and within a day or so saw a substantialy, visually apparent change in corals. or the many, many folks who had dino issues and dosing nitrate helped solve the problem.

it is experimentally validated, IMO.

It's worth thinking about why some N sources and not others, and relatedly (or by analogy) why some carbon sources and not others?
I find it remarkable that the kinds of growth you get from vodka and vinegar dosing water samples are unusual. What I mean is that if you dose enough to grow visible amounts of biomass by dosing vodka or vinegar - you get something that looks weird enough that if a hobbyist had it in their tank, they'd post "what is this stuff?" thread.
My point is that we don't dose glucose or other near-universal carbon sources that can be eaten by everything - probably because the specificity is a good feature, and we'd like to play keep-away from many organisms and grow a tiny subset instead.
Nitrogen dosing has the same issues (maybe). Amino acids are loved by corals - but also people with problem dinos will tell you that their growth is often driven by amino dosing more than any other food additive.
(for background on dinoflagellate use of organic N: see Putting the N in Dinoflagellates and ctrl+F search for aminos. they are quite good at it.)
So maybe that's why we dose the "least useful" N form - Nitrate? We don't want to feed everything.
Or maybe it's the logical fallacy, that we dose NO3 because NO3 is what we measure?

 

[Randy Holmes-Farley]

Does the shuffling of ATP and ADP and vice versa import and export enough PO4 to be notable in the water concentration? Meaning - if I feed some straight carbon source, and the bacteria store up energy as ATP and max out their energy stores - would I see PO4 drop in the water even without the bacteria generating much new biomass?
My guess is no, that the possible pool of internal ATP is small and the cell has other ways to store larger amounts of energy. So I couldn't measure a notable PO4 difference in a water sample between "tired" bacteria and "high energy" bacteria. I wonder if that's true.

That process generally does not itself take up or release phosphate from cells. It happens inside the cells.

 

[Randy Holmes-Farley]

It's worth thinking about why some N sources and not others, and relatedly (or by analogy) why some carbon sources and not others?
I find it remarkable that the kinds of growth you get from vodka and vinegar dosing water samples are unusual. What I mean is that if you dose enough to grow visible amounts of biomass by dosing vodka or vinegar - you get something that looks weird enough that if a hobbyist had it in their tank, they'd post "what is this stuff?" thread.
My point is that we don't dose glucose or other near-universal carbon sources that can be eaten by everything - probably because the specificity is a good feature, and we'd like to play keep-away from many organisms and grow a tiny subset instead.
Nitrogen dosing has the same issues (maybe). Amino acids are loved by corals - but also people with problem dinos will tell you that their growth is often driven by amino dosing more than any other food additive.
(for background on dinoflagellate use of organic N: see Putting the N in Dinoflagellates and ctrl+F search for aminos. they are quite good at it.)
So maybe that's why we dose the "least useful" N form - Nitrate? We don't want to feed everything.
Or maybe it's the logical fallacy, that we dose NO3 because NO3 is what we measure?

Actually, a primary reason I chose acetate (vinegar) is that it is used by every organism, and is widely taken up by all sorts of organisms, including corals, sponges, etc. It is the highest turnover organic compound in many natural environments, including sediments. No need to rely on bacteria, if they are not up to the task for some reason.

 

[Lasse]

Mats form because of vigorous growth and accumulation of ever lengthening filaments and occurs not under nutrient poor conditions.

This is total opposite to hundreds or maybe thousand of different persons observations in working aquaria all around the world - including myself. The triggers for mat forming is nutrient starvation - IMO - there is a overwhelming observed prove for that .

When the mats is formed a new nutrient sink will be available (organic matters and metal bound PO4 together with nitrogen fixation below the mats) and they can continue to grow. When you move them back to a nutrient poor water - without the new nutrient sink available - even my cat can understand that they will disappear and not forms mat - there is no nutrients available to use there and they will use up the hydrocarbons and slowly die. It is not a prove that there is no mat forming trigger when nutrients disappear from the water column - it is a prove that if there is no nutrients in the water column and no nutrients in the substrate they will use up their reserves and slowly die. That is what your experiment have shown not that it is not a mat forming trigger in nutrient starvation in the water column.

So maybe that's why we dose the "least useful" N form - Nitrate? We don't want to feed everything.
Or maybe it's the logical fallacy, that we dose NO3 because NO3 is what we measure?

According to metal bounded PO4 in the substrate - nitrate in the water column will reduce the availability of it but according nutritional needs, your question is very interesting. For the moment - my believe is like the first sentence but there is some observations I have done that will not rule out NH4 totally (according cyanobacteria mat forming but for GHA - NH4 can be the super form )

Sincerely Lasse

 

[taricha]

Actually, a primary reason I chose acetate (vinegar) is that it is used by every organism, and is widely taken up by all sorts of organisms, including corals, sponges, etc. It is the highest turnover organic compound in many natural environments, including sediments. No need to rely on bacteria, if they are not up to the task for some reason.

Faaascinating. Thanks for context.
Interestingly, I think @Dan_P looked at short term (~1 day) bacterial activity in a sample of tank water dosed with vinegar.
At levels on the high end of what people might dose in a tank, bacterial activity seemed to be suppressed.
Probably a pH effect that would be mitigated in a full system with rock etc, but it made me question how universally accessible vinegar really is.

 

[Randy Holmes-Farley]

Faaascinating. Thanks for context.
Interestingly, I think @Dan_P looked at short term (~1 day) bacterial activity in a sample of tank water dosed with vinegar.
At levels on the high end of what people might dose in a tank, bacterial activity seemed to be suppressed.
Probably a pH effect that would be mitigated in a full system with rock etc, but it made me question how universally accessible vinegar really is.

Significance of Acetate as a Microbial Carbon and Energy Source in the Water Column of Gulf of Mexico: Implications for Marine Carbon Cycling

https://agupubs.onlinelibrary.wiley.com/doi/abs/10.1029/2018GB006129#:~:text=Acetate%20is%20a%20key%20intermediate,unconstrained%20in%20the%20pelagic%20ocean.&text=At%20some%20sites%2C%20acetate%20carbon,of%20the%20bacterial%20carbon%20production

.


"At some sites, acetate carbon may have accounted for up to 50.4% of the bacterial carbon production. These results suggest that acetate may serve as an important carbon and energy source for heterotrophic bacteria thus revealing a potentially significant role of acetate for dissolved organic carbon cycling in the ocean."

 

[Lasse]

Meaning - if I feed some straight carbon source, and the bacteria store up energy as ATP and max out their energy stores - would I see PO4 drop in the water even without the bacteria generating much new biomass?

There is bacteria species and strains that is known for this - they are sometimes referred to as Luxury consumer of PO4. As an example some non sulphur purple bacteria is known to do that (if I remember right). Look after terms like "bacterial phosphate removal in wastewater plants"

Sincerely Lasse

 

[flampton]

Actually, a primary reason I chose acetate (vinegar) is that it is used by every organism, and is widely taken up by all sorts of organisms, including corals, sponges, etc. It is the highest turnover organic compound in many natural environments, including sediments. No need to rely on bacteria, if they are not up to the task for some reason.

Actually acetate is not used by every organism. There are various fermenters in the aquarium, obviously a lesser population than aerobic heterotrophs though. That's one of the reasons I'm not a huge fan of acetate as the only carbon source. It's also not especially gluconeogenic, and a lot of organisms will struggle growing on it as their sole carbon source.

And no ethanol doesn't count as a second type of carbon. I have no idea who thought that was a great idea. A very expensive carbon source that has to be converted to acetate and even with the most careful metabolisms will have some acetylaldehyde damage. Enough that matters? Not sure, but why would I want to choose a more expensive option that is more toxic when I'm spending thousands on live stock.

Now what still needs to be established is whether acetate selects a specific population of bacteria that is the most helpful in achieving our specific goals. Which for the majority of people carbon dosing is thought of as N and P sequestration. Now personally I think that mindset needs to change to a more thorough understanding of the overall ecological needs of each aquarium.

 

[flampton]

It's worth thinking about why some N sources and not others, and relatedly (or by analogy) why some carbon sources and not others?
I find it remarkable that the kinds of growth you get from vodka and vinegar dosing water samples are unusual. What I mean is that if you dose enough to grow visible amounts of biomass by dosing vodka or vinegar - you get something that looks weird enough that if a hobbyist had it in their tank, they'd post "what is this stuff?" thread.
My point is that we don't dose glucose or other near-universal carbon sources that can be eaten by everything - probably because the specificity is a good feature, and we'd like to play keep-away from many organisms and grow a tiny subset instead.
Nitrogen dosing has the same issues (maybe). Amino acids are loved by corals - but also people with problem dinos will tell you that their growth is often driven by amino dosing more than any other food additive.
(for background on dinoflagellate use of organic N: see Putting the N in Dinoflagellates and ctrl+F search for aminos. they are quite good at it.)
So maybe that's why we dose the "least useful" N form - Nitrate? We don't want to feed everything.
Or maybe it's the logical fallacy, that we dose NO3 because NO3 is what we measure?

Yes!! You nailed it. People dose nitrate because it's measurable. Then since it was shown to help a bit that became established. I guarantee if you can go back you'll probably see no one even mention the nitrogen flux in an aquarium. I mean I think I'm probably the first person to mention DNRA microbes on this forum. And I mean that's a huge nitrogen recycling system in the ocean.

Speaking of this Hanna released a ulr nitrate test but I would much rather preferred a ulr ammonia test. Though at the levels we're talking about you would need continual monitoring. E.g. a shift in carbon levels lower and ammonia will go up, more carbon added and ammonia levels will go down. You won't see a static basal level of ammonia in an aquarium ever.

 

[Lasse]

I think I'm probably the first person to mention DNRA microbes on this forum

No you are not at all - it has been discussed before but no one have shown them in aquarium. They also need anaerobic condition - and in most ( read bare bottom tanks) - there is very few environment that is anaerobic and have the waterflow that´s needed for transport in and out. IMO most of the NH3/NH4 production - in most tanks - are from aerobic mineralization of organic matter + 2 hours direct transport from the gills of the fish after feeding. In my tank - I´m sure that DNRA exist together with many facultative and obligate anaerobes.

I have advocate for the flux theory in this forum for more than 4 years now and also when it is about the "desert" coral reefs with low left over in the water column.

Then since it was shown to help a bit that became established

Yes and it seems that it is difficult for some to understand this. It works. I notice many years ago that dosing pure nitrate was helpful in 80 % of the cyano outbreaks. I have been very aware of NH4/NH3 importance for nutrient uptake in water - but still readable nitrate levels helped with most outbreak of Cyano. There maybe is another mechanism of biochemical nature and the fight is not about N - it is about P.

Sincerely Lasse

 

[Randy Holmes-Farley]

Actually acetate is not used by every organism. There are various fermenters in the aquarium, obviously a lesser population than aerobic heterotrophs though. That's one of the reasons I'm not a huge fan of acetate as the only carbon source. It's also not especially gluconeogenic, and a lot of organisms will struggle growing on it as their sole carbon source.

And no ethanol doesn't count as a second type of carbon. I have no idea who thought that was a great idea. A very expensive carbon source that has to be converted to acetate and even with the most careful metabolisms will have some acetylaldehyde damage. Enough that matters? Not sure, but why would I want to choose a more expensive option that is more toxic when I'm spending thousands on live stock.

Now what still needs to be established is whether acetate selects a specific population of bacteria that is the most helpful in achieving our specific goals. Which for the majority of people carbon dosing is thought of as N and P sequestration. Now personally I think that mindset needs to change to a more thorough understanding of the overall ecological needs of each aquarium.

Well, I'm a huge fan of both ethanol and acetic acid because they have a very long and established history of success. If your theory says it is bad, well, you might want to examine the theory. lol

As to acetate not being used by every organism, while I admit to not asking every organism in my tank if they use it, it is extremely well established that acetate plays a huge role in the carbon cycle in anaerobic sediments as well as in the water column.

 

[flampton]

No you are not at all - it has been discussed before but no one have shown them in aquarium. They also need anaerobic condition - and in most ( read bare bottom tanks) - there is very few environment that is anaerobic and have the waterflow that´s needed for transport in and out. IMO most of the NH3/NH4 production - in most tanks - are from aerobic mineralization of organic matter + 2 hours direct transport from the gills of the fish after feeding. In my tank - I´m sure that DNRA exist together with many facultative and obligate anaerobes.

I have advocate for the flux theory in this forum for more than 4 years now and also when it is about the "desert" coral reefs with low left over in the water column.


Yes and it seems that it is difficult for some to understand this. It works. I notice many years ago that dosing pure nitrate was helpful in 80 % of the cyano outbreaks. I have been very aware of NH4/NH3 importance for nutrient uptake in water - but still readable nitrate levels helped with most outbreak of Cyano. There maybe is another mechanism of biochemical nature and the fight is not about N - it is about P.

Sincerely Lasse

Obviously almost all production of ammonia is going to be the breakdown of proteins, nucleotides etc. Where did I say that DNRA are mostly responsible for this?

I can tell you that they are in your aquarium. Just like I can tell you there are denitrifiers. I mean DNRA is just another type of denitrification. They live in the same niche...which brings me to

Don't care how bare bottom a tank is there is still anaerobic areas. If you understand the dynamics of biofilms you would know that there are water channels that allow nutrients to spread through the film. However as the biofilm gets thicker the oxygen is stripped by the organisms in the 'top' of the biofilm and the organisms on the bottom are anaerobic. So yeah not buying the bare bottom no anaerobic zone idea. Is it less the a tank with a dsb, I mean obviously but they're there.

Oh and how about on an animal we care about too

Just a 1 minute pubmed search to pull a paper showing anaerobes in this Acroporidae family corals skeleton

Metagenomic, phylogenetic, and functional characterization of predominant endolithic green sulfur bacteria in the coral Isopora palifera - PubMed

We suggest that the endolithic microbial community in coral skeletons is diverse and dynamic and that light and oxygen are two crucial factors for shaping it. This study is the first to demonstrate the ability of nitrogen uptake by specific coral-associated endolithic bacteria and shed light on...

pubmed.ncbi.nlm.nih.gov

 

[flampton]

Well, I'm a huge fan of both ethanol and acetic acid because they have a very long and established history of success. If your theory says it is bad, well, you might want to examine the theory. lol

As to acetate not being used by every organism, while I admit to not asking every organism in my tank if they use it, it is extremely well established that acetate plays a huge role in the carbon cycle in anaerobic sediments as well as in the water column.

Well its not a theory its a hypothesis for one. Two I see that you have left the science field completely and are now just arguing from anecdotal evidence and your belief system.

Yes great plays a role. Can you show me the acetate is the sole carbon source in the water column paper? Thanks I'll wait.

 

[Randy Holmes-Farley]

Well its not a theory its a hypothesis for one. Two I see that you have left the science field completely and are now just arguing from anecdotal evidence and your belief system.

Yes great plays a role. Can you show me the acetate is the sole carbon source in the water column paper? Thanks I'll wait.

My suggestion is to check the attitude when speaking to other scientists here. It will go over better.

As a nationally recognized expert pharmaceutical chemist, I understand biochemistry pretty well. I have spent decades looking into the chemistry of coral reef aquaria, and when I choose to look into dosing of organics more than 10 years ago for a variety of purposes, I read a variety of scientific papers to inform my decision.

The abundance of marine organisms from bacteria to corals to sponges that are known to take up acetate is large, and that knowledge assured me that acetic acid was a decent choice to dose into aquaria. None of us have any idea what fraction of the dosed organics are consumed by bacteria or by other organisms. It would be interesting to know, but it doesn't matter most of the time. It accomplished exactly what I wanted (increased food for filter feeders: I could both see it and over time saw increased growth of sponges), and was a cheaper way to keep nutrients under control than growing macroalgae and its associated electrical costs).

I have read many HUNDREDS OF THOUSANDS of reef anecdotes over the past 25 years. Perhaps more than anyone else (not sure on that). At some point, the weight of the evidence becomes very convincing science. It works. Like knowing that if you go outside your home, there will be air and you do not need to worry about lack of oxygen. I'm certain there has never been a single study of that theory/hypothesis, but the weight of enough anecdotal evidence rules the day.