Do we need Nitrate in a reef tank?

Lasse

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For anyone who can access the original article (I cannot), this article may be of interest to this discussion:


Sincerely Lasse
 

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With all the respect to their efforts to make this hobby more popular I tend to agree. With their resources they can organize experiments in far better way to be scientifically sound. They can make partnership with an University and provide them with topic and resources. There are hundreds of scientifically challenging topics in this hobby and I can see tens of Masters and PhD thesis on them.
Their experiments are to sell products. It's great marketing. But also good enough learning material for hobbyist
 
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I among these that prefer a NO3 concentration between 2 - 5 mg/L. One of the reasons is that if I read NO3 - I know that I have a N reserve in my water but I´m also run Phosphates well over 0.04 in these cases, There is reports that indicate that NO3 levels around 0.2 mg/L and PO4 below 0.03 can cause P defiency and bleaching among some stony corals. With PO4 above 0.04 - the amount of NO3 is not as critical.

Another reason is the fact as someone state - the only organism I know that really need NO3 is the denitrification bacteria. I prefer to see my aquarium as an ecosystem where bacteria may be the most important inhabitants. In a mature aquarium - there is a lot of different environments with there own set up of bacteria. In the part where oxygen is available - oxygen is used by heterotrophic bacteria in their cellular respiration. But there is also parts that are more or less depleted of oxygen (by the same bacteria) where there is still plenty of organic matter to "eat" Many heterotrophic can in these cases switch from aerobic respiration into anaerobic respiration in order to use the the organic resources. What anaerobic respiration really means? It means that oxygen is not available as an electron acceptor in the respiration - some other molecule must be used. Its here the NO3 molecule comes in as the savior in distress. Many bacteria strains can shift between oxygen and NO3 in the respiration cycle. The oxygen will give more energy but NO3 is first runner up in energy yield. The waste from aerobic respiration is as most of us know CO2 and the waste from anaerobic respiration with help of NO3 is N2 (there is a name for this - denitrification) Its important to note that there is many aerobic bacteria that can switch from using oxygen to be anaerobic - using NO3 in the respiration. OK - i want an anaerobic bacteria population that use NO3 i the respiration - why.


The answer is hidden in the fact the second runner up in the respiration race is bacteria strains that only use sulphate as electron acceptors in the respiration. Our aquariums is rich in sulphates - my own sulphate contend you can see below

1681077026321.png


This means that if not NO3 is available - the bacteria strains that use Sulphate as electron acceptor will dominate the anaerobic bacteria population totally. - And whats with that?

The most important "what" is the waste they produce - hydrogen sulphide - the rotten egg smell. Its a very toxic compound and IMO one of most common reasons for sudden fish death in mature system. Yes it can be produced even if NO3 is represented but IMO - measurable concentration of NO3 above 2 mg/L limit the hydrogen sulphide producers biomass because the bacteria that denitrify take up rather much of the available area for bacteria growth.

The bacteria that use sulphate in their respiration seems als to be of certain strains that not can use either O2 or NO3 in their respiration.

There is a third reason too - and its about cyanobacteria. My believe is that sometimes is not the mat building cyanobacteria blooms a result of nitrogen starvation but more a result of available phosphorous sources. Many times it had shown up that very low or total zeroed NO3 concertation result in blooming cyanobacteria mats. I´ll think that there is a link between the anaerobic bacteria population between the mats and the substrate. If no NO3 is present - the hydrogen sulphide producers will dominate. Hydrogen sulphide break the bonds between metals and PO4, hence PO4 will be available for consumption. If I got these mats - I always try to stirr the mats in the evening and add NO3 to the water. I´ll think that stirring the mats is good - even if you not suck them out - the mats is not primary the cyanobacteria - they are - IMO - external resources of mainly sugar produced formed during the daily photosynthesis

This was a long post and probably not exactly right in a scientifically way for a microbiologist but an attempt to explain why I prefer NO3 concentrations between 2 - 5 mg/L

And a attempt to explain why I say yes to the initiala question "Do we need Nitrate in a reef tank?"

Sincerely Lasse

Thank you Lasse for this comprehensive answer. You pointed out three reasons I 've tried to summarise :

1. Under NO3 levels around 0.2 mg/L and PO4 below 0.03 can cause P deficiency and bleaching corals
2. If not NO3 available - the sulfate reducing bacteria (SRB) will dominate over nitrate reducing bacteria (NRB) in the anoxic sandbed and the hydrogen sulphide they produce could kill aquarium inhabitants (threshold about 2 ppm).
3. If no NO3 is present - hydrogen sulphide produced by SRB will break the bonds between metals and PO4, hence PO4 will be available for consumption, resulting in blooming cyanobacteria mats on sand surface.

IMO #2 is very good one -there are many papers on NRB - SRB interactions and ways to reduce SRB with nitrate dosing to prevent corrosion and H2S emissions for practical reasons.

But I have doubts about #1 - not sure if at nitrate levels of .2 ppm and PO4 levels around 0.03 will cause P deficiency. It could be true only if no other nitrogen sources are available. Nitrate reductase is even inhibited in presence of ammonia.

#3 sounds interesting and intriguing theory but not sure this can be the main reason for a cyano bloom. Still nitrogen (but not nitrate) limitation for sure can give advance to cyanobacteria over the others since cyanobacterial cysts are capable of nitrogen fixation.

In relation of the abovementioned is there a positive identification of "red slime" cyanobacteria? I found only few quite opposite opinions.
 

Lasse

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1. Under NO3 levels around 0.2 mg/L and PO4 below 0.03 can cause P deficiency and bleaching corals
No NO3 levels over 0.2 mg/L and PO4 below 0.03 mg/L can cause PO4 starvation and bleaching. As soon as PO4 is over 0.03 - higher NO3 can be tolerated https://www.frontiersin.org/articles/10.3389/fmars.2015.00103/full and https://www.sciencedirect.com/science/article/pii/S0025326X17301601

#3 sounds interesting and intriguing theory but not sure this can be the main reason for a cyano bloom. Still nitrogen (but not nitrate) limitation for sure can give advance to cyanobacteria over the others since cyanobacterial cysts are capable of nitrogen fixation.

Not all cyanobacteria have heterocysts - especially not those benthic ones responsible for the mats. However they can still use bacteria to fixate N2 if an anaerobic zone between the mats and the substrat can be established. I do not know if released PO4 is the main reason but IMO a possibility.

The attached PDF can give some ideas. Unfortunately is written in Swedish but I think that Goggle translate can help to understand where my ideas come from.

Page 44 and forward is interesting and especially from page 51. Chapter 5.7 is interesting in order to understand my ideas according to the mat forming cyanobacteria in saltwater

Sincerely Lasse
 

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No NO3 levels over 0.2 mg/L and PO4 below 0.03 mg/L can cause PO4 starvation and bleaching. As soon as PO4 is over 0.03 - higher NO3 can be tolerated https://www.frontiersin.org/articles/10.3389/fmars.2015.00103/full and https://www.sciencedirect.com/science/article/pii/S0025326X17301601
Yes, as I said it could be true if there is no other nitrogen sources, as it is in the controlled experiment you and others sharing recently. From it: "The ammonium levels found in our mesocosm are very low (< 0.7% of total dissolved inorganic nitrogen) compared to the combined nitrite (~ 10%) and nitrate concentrations (~ 90%). Therefore, the measured NO3− concentrations (combined NO2−/NO3−) represent largely the total dissolved inorganic nitrogen pool that could be accessed by the zooxanthellae in the present experiment."
Still not sure there is a proof in this experiment that nitrate is actually needed in a reef tank, but agree if you have elevated levels of nitrate is probably good idea to have higher levels of phosphate too.
 
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Not all cyanobacteria have heterocysts - especially not those benthic ones responsible for the mats. However they can still use bacteria to fixate N2 if an anaerobic zone between the mats and the substrat can be established. I do not know if released PO4 is the main reason but IMO a possibility.
:) That is why I've asked if there is positive identification of red slime cyano from reef tanks because I've found some say it is Cylindrospermum (which have heterocysts) and others says Phormidium (which doesn't have) - none of them IMO seems to be the correct identification.

Following very good reads can answer the question if cyanobacterial mats can fix N2, but even if you not interested about N2 the first paper is very good in understanding of cyanobacterial mats in the natural reefs and have some very good pictures that could help identifying and understand "our" aquarium cyano.

Brocke et al. 2018. Nitrogen fixation and diversity of benthic cyanobacterial mats on coral reefs in Curaçao

Here more on N2 Fixation by non-heterocystous cyanobacteria
 

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I have seen reports - from aquarists - that in the red mats there can be at least 2 families/species. One of them a spirulina species. Tests with hydrogen peroxid can tell them apart. The spirulina type is less affected than the other. The non spirulina type will be totally be dissolved by H2O2 - the spirulina type not.

I do not know if I right but I have always seen our mat forming (i.e. external deposit of sugar) as belonging to the Oscillatoria group or are very close to that group. No heterocyst in many of them. From your first link - my bold

Microscopic examinations showed 22 cyanobacterial species, all from the order Oscillatoriales.

My thoughts are that both cyanobacteria and dinoflagellates have found their own way to access phosphorous down into the substrate. For cyanobacteria nitrogen is no problem - they have found many ways to fix its own NH4/NH3. Dinoflagellates however - probably need dissolved inorganic N but NO3 free water provoke forming of hydrogen sulphide and the following release a PO4 - make it easier to get PO4 from the sediment pore water

I strongly recommend all who are interested of this to run Chapter 5:7 and 5:8 of the PDF file I attached in post #144 through Google translate. It gives a lot of things to think of

Sincerely Lasse
 
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Randy Holmes-Farley

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So, is the vertical axis a percentage increase, reduction?
If someone could enlighten me on how to convert the horizontal axis to ppm, that would be great, I have no idea, lol


I'm not used to looking at data based on "effect size", but I believe a positive effect size is an increase and a negative effect size is a decrease. hedges d is a complicated unit, involving the difference in means divided by the standard deviation. I've never used such a thing, or even seen it before.

For the x-axis:

1 uM = 17 ppb ammonia, 62 ppb nitrate, or 95 ppb phosphate.
 

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FYI

A way of learning how to convert the concentration unit M (Molar or mole/litre) into weight units is to use the definition of the world Molar. 1 mole of a compound is the atomic mass in g. The hydrogen atom have the atomic mass 1 and one mole of hydrogen is corresponding to 1 g hydrogen atoms. Nitrogen have the atomic mass of 14 - 1 mol nitrogen atoms is therefore 14 g nitrogen. If you take 4 atoms of H with 1 atom of nitrogen - you get NH4 (ammonium)The atomic mass of NH4 is 4*1 +14 (H4+N)= 18. Hence - 1 mol of ammonium is therefore 18 g. M or molar is mol/L with gives us that 1M ammonium content 18 g of ammonium or 18 g/l or 18 000 mg/L or 18 000 000 µg/L. 1mM (milli Molar) NH4 is therefore 18 mg/L - 1µM (micro Molar) is 18µg/l - often referred as 18 ppb. Randy use 17 ppb but I suppose he refer to ammonia gas (NH3) not the ammonia ion (NH4 or ammonium) as I do.

Do the mat on NO3 and PO4 and you will see how the process work. 1 mol NO3 = 14 (N) +3*16 (O) g NO3 -> 62 g NO3 -> 1µM = 62 µg/L (ppb) Atomic mass of PO4 (kind of ) is 95 (31 (P) + 4*16 (O). - 1µM PO4 = 95 µg/L PO4 or around 95 ppb


Sincerely Lasse
 

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Unrelated to current discussion (except it's in this thread.)
Thanks @Timfish for this term "carbonate mud"
Fish as major carbonate mud producers and missing components of the tropical carbonate factory
https://www.pnas.org/doi/10.1073/pnas.1015895108
I've bugged Dan occasionally about how my sand over time shifts from large gains to increasing amounts of fine grained stuff I've called silt.
Most assume it's bacteria/mulm/etc. But the large majority of this cloudy stuff dissolves in a little HCl and a Ca titration on the dissolved stuff gives you close but a little lower than what you'd expect if it were CaCO3.
There are a number of mechanisms to make this stuff, but I didn't know precipitation in fish intestines was one of them.
 

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Unrelated to current discussion (except it's in this thread.)
Thanks @Timfish for this term "carbonate mud"

I've bugged Dan occasionally about how my sand over time shifts from large gains to increasing amounts of fine grained stuff I've called silt.
Most assume it's bacteria/mulm/etc. But the large majority of this cloudy stuff dissolves in a little HCl and a Ca titration on the dissolved stuff gives you close but a little lower than what you'd expect if it were CaCO3.
There are a number of mechanisms to make this stuff, but I didn't know precipitation in fish intestines was one of them.
Thank´s both

Sincerely Lasse
 
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Unrelated to current discussion (except it's in this thread.)
Thanks @Timfish for this term "carbonate mud"

I've bugged Dan occasionally about how my sand over time shifts from large gains to increasing amounts of fine grained stuff I've called silt.
Most assume it's bacteria/mulm/etc. But the large majority of this cloudy stuff dissolves in a little HCl and a Ca titration on the dissolved stuff gives you close but a little lower than what you'd expect if it were CaCO3.
There are a number of mechanisms to make this stuff, but I didn't know precipitation in fish intestines was one of them.
Similar experiment with detritus buildup in my bare bottom tank

 

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Again, commenting on material earlier in thread.
Dosing aminos gives very good results, but requires caution. I started low, and all was fine, but when I have increased the amount to be the only nitrogen source in the amounts that are daily consumed by my system, in about a week or two, it resulted in a severe dino outbreak which then took me months to cure.
This is actually a very common observation. People with dinos often report that aminos really increase the bloom. This is a much stronger correlation than zero NO3 connecting to dinos.
Which brings me to this. I wanted to reiterate this.
[low to zero] Phosphate is the apparent correlate, not available nitrogen. In the several hundred dino cases I have seen, I don't think nitrogen is something to consider as a possible correlate.

... and for the same reason. Just my sense after a large numbers of anecdotes....
Eh its all anecdotal. No one seems to know the answer, and so this is based off of the hundreds of anecdotes I have seen where nitrate doesn't seem to matter as much as phosphate when it comes to dinos.
Dino outbreaks in hobby seem to have strong correlation to people dropping PO4 to zero (especially with GFO - the correlation might go away if you only look at dropping to zero PO4 by other methods, carbon dosing etc. ) and the correlation to zero NO3 is much smaller.

But these aren't universal. There are people who know their tank runs well with zeros, and they aren't concerned with dino advice and just keep on with ULNs.

Years ago (5+), I had dinos, even today I can still see a few cells on GHA under the 'scope. But now trying to do all the things that people - like me - say can "cause" dinos to take over: Lowering PO4 to zero with GFO, and NO3 to zero, amino acids (glutamine and aspartic acid), water changes, trace element dosing etc. Nothing. Keeping those conditons up for a month+ - not a tiny patch of strand of brown anywhere in the tank (still only stray cells on GHA in sump).

So our explanations are obviously incomplete, and our proposed mechanisms are maybe more correlations than causation.
 

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Similar experiment with detritus buildup in my bare bottom tank
That's awesome that you could generate the CaCO3 particle debris in a barebottom tank - no sandbed requred!
 

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Again, commenting on material earlier in thread.

This is actually a very common observation. People with dinos often report that aminos really increase the bloom. This is a much stronger correlation than zero NO3 connecting to dinos.
Which brings me to this. I wanted to reiterate this.


... and for the same reason. Just my sense after a large numbers of anecdotes....

Dino outbreaks in hobby seem to have strong correlation to people dropping PO4 to zero (especially with GFO - the correlation might go away if you only look at dropping to zero PO4 by other methods, carbon dosing etc. ) and the correlation to zero NO3 is much smaller.

But these aren't universal. There are people who know their tank runs well with zeros, and they aren't concerned with dino advice and just keep on with ULNs.

Years ago (5+), I had dinos, even today I can still see a few cells on GHA under the 'scope. But now trying to do all the things that people - like me - say can "cause" dinos to take over: Lowering PO4 to zero with GFO, and NO3 to zero, amino acids (glutamine and aspartic acid), water changes, trace element dosing etc. Nothing. Keeping those conditons up for a month+ - not a tiny patch of strand of brown anywhere in the tank (still only stray cells on GHA in sump).

So our explanations are obviously incomplete, and our proposed mechanisms are maybe more correlations than causation.

If the goal of having available nutrients is to drive competition for the dinos, then presumably one would optimally use forms that dinos could not themselves use, but the competitors could, or at the very least, not add things dinos can use but comeptitors cannot.

Amino acids seem preferred by dinos but not preferred by algae.


"
Curiously, different blooming populations of dinoflagellates were found to have high uptake rates for urea and/or amino acids, and these rates were always higher than the rates for NO−3
uptake (Kudela and Cochlan, 2000; Fan et al., 2003; Collos et al., 2007). In L. polyedrum, the urea uptake rate was also about 2 times more than that of NH+4, even if environmental urea concentrations were less than NH+4
(Kudela and Cochlan, 2000). Taken together, these observations suggest that dinoflagellates possess a full suite of transporters for inorganic N and organic N forms, that they have the biochemical means to assimilate these N forms, and that they show a great physiological plasticity in response to external N types and concentrations."



"Attempts to measure the uptake of amino acids by natural algal populations have failed to show any significant levels of uptake by microalgae. "

"Microalgae can take up amino acids from natural concentrations found in seawater"

"if the microalgal utilization of DFAA, and of other as yet unconsidered components of DON, were shown to be significant, either in terms of nitrogen or carbon input, it would have considerable repercussions on our conception of the ecology of phytoplankton."
 
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That's awesome that you could generate the CaCO3 particle debris in a barebottom tank - no sandbed requred!
Yes, I think it comes from the rocks mostly. All the micro creatures that live in there and digging tunnels or carving the surface are main contributors to this IMO . But precipitation can not be excluded too. I was dosing two parts by the time.
 

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and carbon dosing 10 ml daily,
IMO - Here is your explanation why you had more organic detritus than your feeding should cause. IMO - organic carbon is mostly the limited factor for heterotrophic bacteria growth in our aquariums. With adding DOC - as you did - its not limited any longer and bacteria could growth on surplus N and P. Much of your organic detritus ought to be just bacteria mulm.

Sincerely Lasse
 

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or at the very least, not add things dinos can use but comeptitors cannot.

Amino acids seem preferred by dinos but not preferred by algae.
This effect seems quite solid. And is one area of dinos where the published lit lines up perfectly with what we see in the hobby.
I've also seen this in action on a small scale. A tiny test tank had a mix of Macros, GHA and dinos (zero detectable NO3) adding amino acid product as only N input, the GHA receded and the dinos overran things.


With adding DOC - as you did - its not limited any longer and bacteria could growth on surplus N and P. Much of your organic detritus ought to be just bacteria mulm.
...and of course algae of all sorts can generate particulate organic detritus as it dies or is eaten - the degraded tissue's mass should mostly be from captured CO2. Represents another way to get debris in excess mass of food input.
 

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Sand dissolves. Over the decades, I have to replace it. The pH down in the depths can be in the 6s if you are deep enough to have anoxic zones. A lot of smaller particles come from larger sand.

If a tank has enough building blocks for the "competition," then there is enough for dinos. Ugly tanks has always been about sterilie/fresh locations for the faster growing stuff to colonize first... which is dinos and cyano. Think dandelions and crabgrass on a fresh dirt lawn before grass can take over - this is not a "nutrient" problem either. There is nothing more to it than this, unless you can growth limit dinos with higher no3 and po4 levels.
 
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...and of course algae of all sorts can generate particulate organic detritus as it dies or is eaten - the degraded tissue's mass should mostly be from captured CO2. Represents another way to get debris in excess mass of food input.
Agree on that. At least in my case most of the detritus apart of calcium carbonate was actually semi digested algae, it was clearly seen under microscope. Cellulose is hard to digest in aerobic conditions. That’s why I’m having sandbed now and decomposition of cellulose is much faster. my sandshifting goby does great job to burry all the nasty stuff deep enough for anaerobes to take over.
 

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