When and why did dosing nitrates start?

[Dan_P]

Exactly - they have to seek it in another place when the left over in the water column gone away. In sediments or in organic matter. 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 . The mats - the secret is mostly hydrocarbons from the photosynthesis - giving carbon to the bacteria working below the mats and to the cyanobacteria IMO. The mat forming bacteria is always present - the mats not. If you can repeat your test with PO4 and minerals in the water but only get mat forming when you ad amino acids - you have got something - but use F/2 solution this time (with and without inorganic N)


A good sign that it takes time

Sincerely Lasse

I don’t recall ever reading that mats form in response to a lack of food. Rather mats form as a consequence of good conditions. High density, fast growing microorganisms need a heavy supply of nitrogen and phosphate, removal of oxygen and supply of CO2. Microorganisms typically disperse when nutrients become scarce.

 

[ichthyoid]

Thank you Lasse

 

[Lasse]

I don’t recall ever reading that mats form in response to a lack of food.

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

 

[taricha]

one minor quibble. The O2 is being reduced to water. The CO2 is from the oxidation of the carbon of the glutamine.

ooh. I missed that. O2 disappearing in ways other than by hooking up with Carbon.

 

[flampton]

ooh. I missed that. O2 disappearing in ways other than by hooking up with Carbon.

Yup, take a look back in one of my posts but the electron transport chain/oxidative phosphorylation uses the reducing equivalents generated prior as the glutamine was being oxidized. Basically what would happen aerobically is the below-

to keep everything a little bit better balanced we're going to use two l-glutamate molecules

1) 2 glutamate + 2 NAD+ --> 2 a-ketoglutarate + 2 ammonia + 2 NADH

The a-ketoglutarate then enters the TCA cycle

2) 2 a-ketoglutarate + 14 NAD+ 5 FADH+ 5 GDP + 5 Pi --> 10 CO2 + 14 NADH + 5 FADH2+ 5 GTP

Then the above NADH and FADH2 enters the etc/ox phos

3) 16 NADH + 5 FADH2 + 10.5 O2 + 58 ADP + 58 Pi --> 16 NAD+ 5 FADH+ 10.5 H2O + 58 ATP

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 This step is also known as respiration and that is why organisms that respire can grow faster than organisms who don't have that capability.

 

[flampton]

Sorry just realized it was glutamine not glutamate.

So one extra step
Glutamine --> glutamate + ammonia

So no extra energy but a extra ammonia. 2 ammonia molecules per glutamine.

 

[flampton]

Now let's look @taricha experiment. You'll see that he used 0.4 mM glutamine. Glutamine weighs 146 g/mol. So he added 58.4 mg/l glutamine. From this about 7mg/l ammonia was exported into the water. Of this it is 5.8mg/L nitrogen atoms. The nitrogen make up around 21% of the glutamine. So let's take that 58.4 and multiply by 21%
We get 12.6 mg total nitrogen in the glutamine.

So what this means is that 46% of the ammonia was released into the water and 54% was used by the bacteria.

So overall conclusion would be ~50% of the nitrogen went towards what we could refer to as ammonia dosing. While 50% of the nitrogen went towards carbon dosing support.

Meaning in this experiment l-glutamine would decrease the effect of carbon dosing by 50% on nitrogen. It however would not effect the amount of phosphate removal.

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? No idea but is again a way to conserve nitrogen while removing phosphates. It could even be cheaper than dosing acetate and ammonium chloride

Also the reverse can be done and phosphorylated sugars or solubilized phospholipids could be added. In this case it will likely stay hypothetical because would be cheaper to add acetate and phosphate.

 

[Seabiscuit]

Keep in mind that I am not a novice. I have maintained a reef for over two decades and have reset my tank multiple times with similar results, BUT to the point of this thread I ALWAYS test zero on nitrates . . . and I use the highest quality tests and multiple variations . . . I have never found nitrates. Obviously, my system has nitrate that occurs. My question is: Does having trace nitrate in your system improve coral health, and if so what proof is there.

Love those shots! Nice!

 

[Sallstrom]

Great discussions!

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).
I have my own ideas and have tried some of them but not in any scientific way, and not always with clear results

 

[Lasse]

Love those shots! Nice!

The funny thing is that there is a lot of tanks running in NO3 levels 2 - 5 mg/L (and even higher) that looks like that too. There is - IMO - fewer problems to run close to zero in established tanks like the one in the pictures without getting serve cyanobacteria problems compared with newly established tanks. IMO - it is much easier to establish and start a new tank with a readable NO3 (and PO4) concentrations. IMO - if anyone want to hit 0 in nutrients - do that after a decent biomass of corals are established - this will take some time and if it are the reefers better skills or the domination of corals that´s the most important factor - I can only guess

have reset my tank multiple times with similar results,

May I ask the reasons for the resets and if you run any "method" like zeovit? Any adding of amino acids or other masked nutrient products?

@flampton In your assumption - you see the uptake of amino acids solely as N and C source for further processing (or release). In my mind - one of the reasons for cells fast uptake of these organic N and C containing compounds are that many of them can be used directly in the protein synthesis as a complete part of the produced proteins. As an example - In human there is nearly half of the amino acids we need thats has to be imported into the cells as a complete amino acid for the protein synthesis because we can´t produce them by ourselves from other amino acids or of other compounds.

With other words - you forget the building of biomass in your calculation and for ectothermic organisms it usually are between 20 - 25 % of the intake of nutrients

Sincerely Lasse

 

[Randy Holmes-Farley]

Again the question was why dose nitrates? I say it is not based on any available evidence as usable nitrogen for most everything we care about is in ammonia or amine compounds. Even the organisms that can utilize nitrate for tissue building must convert it back to ammonia first, which is metabolically expensive. Want your corals and their Symbiodinium happy, well give them their ammonia

Certainly it is.

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.

 

[Lasse]

Or cyano problem.

Sincerely Lasse

 

[2Sunny]

Really Nice system!
I think people say zero nitrates isn't enough, because it's easier to talk about the Nitrate number than the flow of N through the system. The better question might be something like "What's your Nitrate and How many mg/L of protein per day go into your tank?"
Also when I see great happy coral systems like yours that have low N, they frequently have a lot of small active fish (chromis & anthias in your case) that eat many times per day. So again, you show "zero" nitrate, but have constant N flow through the system.

Thank you and thanks to everyone that has made this a great and interesting discussion.

This was my assumption all along. I also assumed that such would be the goal of most reefers.

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

Wait . . . you own a Kia??? I always figured you for a 1980's vintage SAAB driver. How disappointing ;Happy

I think we started to test dosing KNO3 around 2010 at work(small public aquarium). We started taking better notes in 2013, so from that year I know we added both nitrate and phosphate when needed
It was a good discussion on a Swedish forum ~10 years ago, with @Lasse as one of the leading persons, and since then we’ve dosed both NO3 and PO4.
Nowadays we use nitrate, ammonium and amino acids for adding N.

Seriously, though I think this and Lasse's comments have answered my question. If I understand the discussion properly, a decade ago folks noticed either a sudden increase in problem algae in low nutrient systems and/or diminished animal health because aquariums had become "too clean" so at some point a discussion arose as to how to solve the problem of too low nutrients, and I get it that folks who enjoy careful monitoring of parameters chose to dose precise amounts of N in one form or another.


However . . . my primary takeaway from this discussion is: 1) Lasse should switch to pasta since potatoes keep disappearing (and buy a Volvo while he's at it) and 2) me personally, if I feel nutrients are too low, then I will add some fish.


Oh, and Lasse you asked why I "reset" my aquarium. Mostly it happened because my SPS became so thick and overgrown that no amount of thinning would allow me to keep the reef looking nice and staying healthy, plus I rather enjoy the "do over" process as I get to see tiny frags turn into large healthy reefs. I'm currently about 1 year into my latest reset. Some faster growing colonies are already nicely grown, but I have another year or two for the slow growers to fill in . . .

 

[Lasse]

(and buy a Volvo while he's at it)

I drive a SAAB once (1970) - it end up 200 meters out in a field - never more

I´m from a Volvo family - my father work in the engine factory for more than 40 years, my brother wife and their two sons work there still. To around 1998 I always drive a 10 - 15 years old 245 (large family) but when I released that I could lend money and by a new small car and have monthly cost that was lower than I had for my 245 that was 15 years old - I change path and buy a small Hyundai. Since around 2006 - I have always have had a KIA picanto - and I promise you - I´ll never go back I get nearly 3 Picanto for the prize of one Volvo . Yes - I have Jebao pumps too

Lasse should switch to pasta since potatoes keep disappearing

Already done that - I dose NH4 for the moment

Sincerely Lasse

 

[Midrats]

I predict trickle filters becoming chic again to avoid the hassle of having to dose nitrate. All the kids will have to have one. Albert Thiel RIP would have raked in the greenbacks selling bio balls!

 

[flampton]

Certainly it is.

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.

I can agree that nitrates probably helped however what I'm saying is that the available evidence does not suggest nitrates would be a better option than using the preferred ammonia until you see nitrates. Plus...

My bet is that what is happening by dosing nitrates is that you give a nice boost to the DNRA population. So for some reason no one talks about the dissimilatory nitrate reduction to ammonia that's occurring in our fish tanks. The DNRA microbes utilize nitrate as the electron acceptor but only reduce it to ammonia. What this means is if you have nitrate in your tank you'll always have a little ammonia in your tank

Oh and I'll write more up on this but the DNRA population helps control the hydrogen sulfide production as well! So you want these guys (HT to @Lasse for sharing that nitrates help control H2S production)

 

[flampton]

The funny thing is that there is a lot of tanks running in NO3 levels 2 - 5 mg/L (and even higher) that looks like that too. There is - IMO - fewer problems to run close to zero in established tanks like the one in the pictures without getting serve cyanobacteria problems compared with newly established tanks. IMO - it is much easier to establish and start a new tank with a readable NO3 (and PO4) concentrations. IMO - if anyone want to hit 0 in nutrients - do that after a decent biomass of corals are established - this will take some time and if it are the reefers better skills or the domination of corals that´s the most important factor - I can only guess


May I ask the reasons for the resets and if you run any "method" like zeovit? Any adding of amino acids or other masked nutrient products?

@flampton In your assumption - you see the uptake of amino acids solely as N and C source for further processing (or release). In my mind - one of the reasons for cells fast uptake of these organic N and C containing compounds are that many of them can be used directly in the protein synthesis as a complete part of the produced proteins. As an example - In human there is nearly half of the amino acids we need thats has to be imported into the cells as a complete amino acid for the protein synthesis because we can´t produce them by ourselves from other amino acids or of other compounds.

With other words - you forget the building of biomass in your calculation and for ectothermic organisms it usually are between 20 - 25 % of the intake of nutrients

Sincerely Lasse

I didn't assume anything. The fact is that very little of any amino you add will be used as that amino. So the majority of the bacteria in your water column are prototrophs. They make all their amino acids, vitamins, etc... They are going to have first shot at any added amino. Depending on their current needs the carbon part could become structure or energy. The nitrogen will also become either structure or released.

So like I said earlier in this thread if I feed my heterotrophic bacteria ammonia and carbon I'm getting something vastly more nutritious as an end result. All the aminos, lipids, vitamins, nucleotides, carbohydrates etc.... And they sequester phosphate away from the autotrophs!!

 

[flampton]

Great discussions!

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).
I have my own ideas and have tried some of them but not in any scientific way, and not always with clear results

Hi @Sallstrom was wondering if you're asking me this question?

 

[Sallstrom]

Hi @Sallstrom was wondering if you're asking me this question?

The question was to all of you in this discussion. So yes, I would be glad to hear what you think

 

[flampton]

The question was to all of you in this discussion. So yes, I would be glad to hear what you think

I have my thoughts on this...the problem is that I'm currently formulating it and I don't want to reveal a work in progress. More research is required.

Rant time...
Now I will tell you what I see out there in relation to controlling N and P, basically ALL the companies make products with no or very little thought to the actual true biological mechanisms or they hide this knowledge from the general public as a trade secret. And some companies actual market things with implausible mechanisms and I do not know if they know this in house and just don't care or what.

Anyways I know companies want to make money but I wish it wasn't so wacky wild west out there(and hey I understand that if you can get away with selling vinegar and denatured alcohol for 100x profit you'll not change any time soon).

So angel investors out there if your looking to help me start a reef company based on biologically sound products, well call me.