Reflects before you begin dosing carbohydrates

[Belgian Anthias]

Originally Posted by bertoni

I am not sure what your point is. Whether nitrate consumption is "optimal" or not seems irrelevant to me. We are not discussing methods of maximizing growth.

As I have said repeatedly, funding for aquarium research is effectively zero, so we likely will never get high-quality data. You are postulating that a widely-used practice that has a long track record of safety. The burden of proof is on you, in my opinion.

I wanted to point out that the growth rate when using ammonia is not maintained when using nitrate. For commercial farming this is an important issue.

Not all research is funded.
Not all research is done for commercial purposes.
A lot of research has been done for master thesis’s.
Marine aquaculture is booming business.
A lot of research is done for and in public aquaria.
There has been a lot of funded research recently on closed marine systems and its biofilters. AOA , ANAMMOX, COMAMMOX was detected due to extend research on aquarium bio systems.


All articles which are used as references in this threat are based on research on which is referred to in the article. How and why!

Managing a commercial ZMAS based on carbon dosing is only possible due to previous research. It is not on experimental base as is carbohydrate dosing in a marine aquarium on the present day.

No home aquarium is the same but bio-chemical principles in seawater are the same in all closed marine systems.

No biofilm will be exactly the same but the principles on how the bacteria connect and communicate are for all biofilm the same.

We did a lot of research to find out how and why? if you want to fund our research and support our wiki Makazi Baharini, donations are possible! Link available on demand! Our research is accessible for free.

[Belgian Anthias]

Originally Posted by bertoni

You are welcome to your opinion. I don't find the evidence you have presented to be convincing. We will have to continue to disagree.

Is it disagreement or deniel?

No evidence has been provided as far that the information is not correct.

[bertoni]

Given the number of tanks that are successful with carbon dosing, with no signs of trouble, I don't think I'm in denial. I've never heard of even one tank going through the type of collapse that you are positing.

[Belgian Anthias]

When one messes with nature assuming nothing can go wrong one may expect disaster.
It is good to know what is happening while manipulating nature to prevent something may go wrong and what to do when it expectedly or unexpectedly goes wrong. That is why we do research. To know how nature works to be able NOT to mess with it but use nature into our favour.

Before adding supplemental organic carbon to a live support system some questions needed answers.

During carbohydrate dosing:
What happens to the carrying capacity of the system?
What happens to the established biological balance ?
What happens to the nitrification and denitrification capacity installed in the system?
What changes within live rock during carbohydrate dosing? In DSB and other biofilters?

And most important:
What may happen to an aquarium system in which the nitrate level is kept in balance using carbohydrates when for some reason dosing is stopped or interrupted?

Are the imposed changes of no significance? Can the changes have consequences? Can they create a dangerous situation or even be disastrous?

Are the taken risks accountable for the results

How much carbohydrates must be used to lower the nitrate level?
A simple question when all parameters are taken into account. How to determine the parameters needed?

We where able to answer all the above questions with the exception of the last one.

We know that keeping on a high C:N ratio will shift the carrying capacity to heterotropic ammonia reduction.
When organic carbon doses are based on the nitrate level one has no control over the C:N ratio and in a situation where the shift is completed the carrying capacity of the system will become dependable of the carbohydrate supplements. For me such situation is a threat for the continuity of the system and is best avoided ?

My conclusion:

As the C:N ratio in the aquarium is not controlled by the way dosing is done for the moment, carbohydrate dosing is not without risk.

Compared with the possible influence of the nitrate level on the system, the influence on the system by recycling some of this nitrate by carbon dosing is huge and may create a most undesirable situation.
Is it worth the drastic changes induced by carbohydrate supplements to the installed biological balance and carrying capacity of the system just for reducing some nitrate?

[Belgian Anthias]

Originally Posted by Dan_P

I wondered about this. This is probably the explanation for the delay in nitrate reduction when dosing is started.



A lot of carbon goes into energy requirements, production of CO2. I think something like more than 50% of the carbon consumed goes to CO2. This is one factor preventing aquaria from turning into sewage plants.

Not only the fact that ammonia is used first is responsible for the delay. Bacteria coming into contact with a new environment go in there Lag phase ( Bacteria population growth has four phases: Lag, Log, steady, dying ) This is the phase for the bacteria to adapt to the new environment by adjusting there internal pathways, building up tools and modify processing installations, to what is coming and do necessary repairs. The population thus not grow. This phase may take some time, Form one hour to several days. Than the cell will grow to double it's size and the Log phase is started http://www.baharini.eu/baharini/doku...3%ABn#de_groei
As nothing happens to the nitrate level during this Lag phase and during ammonia take up in the Log Phase, users who are not patient enough my add more carbohydrates creating a bacterial bloom in the Log phase. As the growth is exponential also the dying phase will be exponential as the cells of the same generation will start the dying phase at the same time. When the growth is explosive also the dying phase will be explosive which causes a lot of stress within a closed live support system. Bacterial bloom is not that innocent as one often can read due to explosive consumption of oxygen and building materials and following explosive decay of biomass.

A lot of commercial organic carbon additives contain bacteria cultures. Even when these cultures would come to live I am pretty shore that the Lag phase may take a lot of time, more time as the installed bacteria may need.

If all organic matter would be oxidized during re-mineralisation, all organic carbon would be transformed into CO2 http://www.baharini.eu/baharini/doku...emineralisatie That is why nature let heterotropic bacteria grow +-5X faster as autotrophic bacteria. if it was the opposite no organic carbon would be available.
As a closed aquarium system is not really closed as it is still in contact with the atmosphere the side effects of messing with the carbon balance will not provoce much problems. In a system where bioreactors are used this is a complete other story.

A lot of people add products to aquaria of which they do not know what the side effects may or will be?

[nogascans]

Very interesting and openly debated thread which I find refreshing. I don't have much scientific input to add (took a high paying job and never used my marine science schooling).

An observation of my experience and to see if anything may relate. In advance I am sorry if there is nothing here to add to discussion.

I recently (Late December) put together a new tank (Red Sea Reefer 750XXL), and moved over a week all my livestock, rock (new sand) from existing 90 bow-front to new tank.

Older tank was sump-less, and utilized a Bio-Pellet reactor with HOB skimmer which has been in use for last 4+ years. Tank was a mixed/heavy SPS tank, and fish health was always good, coral growth/coloration average with growth and good/not great colors. Generic 6-Bulb T5HO for lighting. Older tank was never an algae magnet, and never saw any HA, Bryopsis, or Dinoflagellates during tank life.

New tank has a sump and I transferred Chaetomorpha/Pods from established system to new tank. Did not run a normal cycle, but instead relied on existing Bio-mass in substrate (130 lb. of existing encrusted rock). Using a new R/O Classic 202S, 3100 GPH return pump, and pair of MP40QD for filtration and flow. I used all new water (Fritz) at same parameters as old tank for Alk, Ca, and Mg (8.5, 430, 1380). ATI 8 bulb T5HO for lighting in tank and LED light for Sump.

I have monitored for ammonia and never saw any measurable signs. I had no noticeable negative effects to corals, and continued encrusting was noticed from day 1. I used PAR meter to adjust lighting to previous tank lighting to try and eliminate any coral shock. I did note immediate improvement in coloration on all SPS, along with some initial lightening of LPS present. Chaeto has grown at a steady state. I have been fighting very low Nitrates <1 (Red Sea) and phosphates <.17ppb (Hanna ULR).

I did experience the "normal" diatom bloom with new rock (was washed before use. I have been fighting a small Dino outbreak at week 3, and not sure if part of a mini-cycle or some of the imbalance issues discussed here with reduction or loss of colonized nitrifying bacteria cultures. Not a terrible by any means outbreak, and use of filter bags has eliminated most of signs without lighting reductions.

Other than a swift and complete stop to organic dosing, I just thought my case was worth mentioning and that it might show some light on the subject being discussed.

Sincerely,

David

[bertoni]

Data always are interesting.

Did you disconnect the bio-pellet reactor for the move? I was a bit unsure on that point.

[Belgian Anthias]

What kind of bio-pellets where used?
Do they leak( dose) organic carbon into the water column or is the carbon available only for the bacteria growing on it?

[nogascans]

Originally Posted by bertoni

Data always are interesting.

Did you disconnect the bio-pellet reactor for the move? I was a bit unsure on that point.

I left it running in old tank until I moved rock (bucket flushed with new water and into new tank). I did not transfer BP to new tank.

Sincerely,

David

[nogascans]

Originally Posted by Belgian Anthias

What kind of bio-pellets where used?
Do they leak( dose) organic carbon into the water column or is the carbon available only for the bacteria growing on it?

I used the BRS brand, and have been for several years. I liked the way they broke down and that made it easier to get correct flow. Active bacteria was visible on media as well as large population of tube worms, etc.:

https://www.bulkreefsupply.com/brs-bulk-biopellets.html

Sincerely,

David

[bertoni]

Thank you for the clarification!

[Belgian Anthias]

Biopellets are made of PHA but there are a lot of different monomers possible containing different quantities of carbon and hydrogen . No specs available for BRS bulk pellets about carbon content but they claim to be 100% pure PHA. Biologically cultivated PHA may be +- 70% of the total dry mass. To get pure PHA in most cases solvents are used. To let it stick together to make pellets some additives must be added.
It is also claimed that:

Nitrate laden Bacteria is removed via the protein skimmer
Bacteria directly converts nitrite into nitrous oxide, so nitrate is never produced
Bacteria consumes nitrates and processes them down into nitrogen gas
Nitrate laden Bacteria is consumed by tank inhabitants like corals and sponges

All needs some explanation!



It is not that difficult to cultivate PHA


Used in a " tumbling reactor" , a moving bed reactor, erosion is high and particles of the pellets will spread into the water column. The PHA becomes part of TOC. The dose organic carbon released and available in the water column is difficult to determine, is not controlled, This way a high C:N ratio is possible in the water column.

When the erosion is limited by preventing tumbling the reactor may clog due to the growth.. The supply to the reactor can be managed by the flow rate which gives some control.
As in the reactor enough carbon is available the limiting factor will be the available other building materials as nitrogen and phosphate. As building materials present in the reactor may be used up, most bacteria and archaea may start to make there own PHA which will be used when they spread in the aquarium.

An unexpected bacterial bloom! Can it be caused by bacteria who have started to use there stored PHA.?

[Belgian Anthias]

Before starting adding supplements to an aquarium on should have some basic knowledge about how bacteria grow and what are the main differences between heterotrophs and autotrophs. A lot of commercial organic carbon additives claim to contain dried bacteria and building materials. Why dried bacteria?
Starters often use supplements of dried nitrifiers in the hope to speed up the nitrifying capacity to support the carrying capacity. Why the need to speed it up?
One must be aware that all these efforts may be for nothing when in a later phase organic carbon will be dosed to control nitrate.
Why not start up the system dosing organic carbon preparations containing bacteria, minerals and all building materials, ?
I found some interesting basic information on the internet which can be used to start the discussion. http://neospark.com/images/AutotropVsHeterotrop.pdf

[Tripod1404]

As a side note, you can (atleast in certain tanks) achieve carbon dosing-like effects by dosing amino acids.

For example, I dose amino acids to reduce nitrate levels and decrease dosing if I want it to increase nitrate. It actually sometimes cause my system to go very nitrate limited and I have to add a mixture of potassium and sodium nitrate to increase nitrate. During this period I have to stop dosing amino acids otherwise nitrate is quickly consumed. I read people who dose amino acids sometimes made similar observations.

Considering bulkier amino acids with large side chains actually have more carbon than nitrogen, it is not that surprising. For example tyrosine has 9 carbons and 1 nitrogen, tryptophan has 11 carbons and 2 nitrogen, as a comparison acetic acid and methanol has 1 carbon and ethanol has 2 carbons.

The only main difference for dosing amino acids is the technical difficulty as you would need to turn of the skimmer for 30 mins to 1 hour, otherwise the skimmer would remove most of what is dosed (at least hydrophobic amino acids).


Therefore I prefer the term "carbon" dosing rather than "carbohydrate" dosing. Amino acids are not carbohydrates but you can achieve the same outcome. From a pure organic chemistry perspective, neither are ethanol or methanol are carbohydrates (FDA sometimes classified ethanol as a carbohydrate but it is scientifically inaccurate), they are alcohols and they dont have the correct hydrogen/oxygen ratio to be classified as carbohydrates. Only alcohols that can be considered as carbohydrates are sugar-alcohols (polyols) like glycerol, sorbitol, manitol, etc. Most carbohydrates are polyhydroxyl derivatives of ketones and aldehydes and that even makes acetic/ethanoic acid a questionable carbohydrate since it is hard to argue it is polyhydroxylated (more like monohydroxylated).

[bertoni]

I'm not sure why amino acids would work better for nitrate reduction, since they are a nitrogen source. I agree that the amount of carbon per unit of nitrate is significant, though. Maybe some organism is limited in growth by a missing amino acid. I'd stick with vinegar, ethanol, or bio-pellets for nitrate reduction, at least for a first attempt, but amino acid dosing can be helpful, at least in theory, depending on what's in the food going into the tank. It's interesting that you get good results. Maybe we should encourage others to give it a shot if they're stuck. More puzzles.

[Dan_P]

Originally Posted by Tripod1404

The only main difference for dosing amino acids is the technical difficulty as you would need to turn of the skimmer for 30 mins to 1 hour, otherwise the skimmer would remove most of what is dosed (at least hydrophobic amino acids).

Usually hours are required for the entire tank volume to be treated by the skimmer and that says nothing about the recovery rate, just passing through the skimmer. Recovery, or in this case loss of the amino acid, by skimming will likely be slow, more so if the skimmer is making dry foam. Keep the skimmer on, the bacteria will harvest the amino acid faster than any skimmer could.

[Tripod1404]

Originally Posted by bertoni

I'm not sure why amino acids would work better for nitrate reduction, since they are a nitrogen source. I agree that the amount of carbon per unit of nitrate is significant, though. Maybe some organism is limited in growth by a missing amino acid. I'd stick with vinegar, ethanol, or bio-pellets for nitrate reduction, at least for a first attempt, but amino acid dosing can be helpful, at least in theory, depending on what's in the food going into the tank. It's interesting that you get good results. Maybe we should encourage others to give it a shot if they're stuck. More puzzles.

Yeah that is the question. I actually accidentally realized this. I was trying to raise nitrates by dosing amino acids, but more I dosed more it reduced ( I actually dropped to undetectable NO3 levels on salifert kit). Than I decided something was not right, I stopped dosing aminos and nitrate slowly started to increase. After that I started to use a mixture KNO3 and NaNO3 to increase nitrate. After that I realized dosing aminos increase nitrate consumption. Actually to a point that if you dose them together, nearly half of dosed nitrate disappears the next day.

While thinking about it I realized that at cellular level, it is quite common for cells to convert glucogenic amino acids to glucose and ketogenic amino acids to fatty acids. Actually this is how the metabolisms of carnivores work, their diets are very poor in carbohydrates (and fats depending on source) they basically convert amino acids to sugars and fats.

Now we dont know the exact amino acids composition of the amino acid additives. But if they have a large content of large amino acids like tyrosine, tryptophan, etc , it would mean they contain a lot more carbon than nitrogen. Plus these large amino acids are not very abundant in proteins, generally smaller amino acids are a lot more common. Below is a link showing the average amino acid content of E.coli proteins;

http://kirschner.med.harvard.edu/fil...upernatant.pdf

4 most common amino acids found in E.coli proteins is glutamate(5C,1N), asparatate(4C,1N), alanine (3C,1N) and glycine (2C,1N) . Aside from this I know glycine is commonly produced by cells as a osmoregulator, so it might be a lot more common outside of proteins. And 4 least common amino acids are tryptophan (11C, 2N) histidine (6C 3N), methionine (5C, 1N) and tyrosine (9C, 1N).

So if the amino acid supplement contained a lot of large amino acids, it would force cells break down the large amino acids and to absorb more nitrogen from the environment, in order to produce more of the smaller, more common amino acids. Like from a 11C tryptophan has enough carbon to built 5.5 glycine. But it only contain 2N, so to built 5.5 glycines a cell would need to pick up 3.5 molecules of nitrate. So it would cause nitrate uptake.

Does it provide any benefit over regular vinegar dosing, I doubt this. The only possible benefit might be uptake of some of the amino acids by corals and other inverts (especially clams). Plus it is hard to overdose it and excess can be removed by the skimmer. But it is far more expensive and can get contaminated.

[Tripod1404]

Originally Posted by Dan_P

Usually hours are required for the entire tank volume to be treated by the skimmer and that says nothing about the recovery rate, just passing through the skimmer. Recovery, or in this case loss of the amino acid, by skimming will likely be slow, more so if the skimmer is making dry foam. Keep the skimmer on, the bacteria will harvest the amino acid faster than any skimmer could.

I use a over sized skimmer, so I am very cautious about how much skimming is going on. Aside from that it causes the skimmer to foam like crazy and overflow, if I dont turn it off.

[Belgian Anthias]

How difficult and expensive one can make a simple task as removing non toxic nitrate?

Do we want to remove the nitrogen or assimilate and recycle it?
Depending of the daily quantity nitrate to remove.
Do we want to keep the present nitrification capacity or shift the ammonium reduction towards organic carbon availability and bacterial growth?
Labor-intensive or normal maintenance practice?
Are all side effects known and how they are prevented?

Removing nitrate? Just controlling the nitrate level? Or do we want to have full control over the daily nitrate removal rate and close the nitrogen cycle?

For removing nitrate from the system we have to reduce it to nitrogen gas.
Controlling the nitrate level is possible both ways, denitrification or assimilation, depending of how much nitrate has to be reduced on a daily base. ( preventing bacterial slime, high C:N ratio's)
For full control over the daily nitrate removal rate one needs a bioreactor and have control over the HRT, the flow rate.
Simultane nitrification and mixotropic denitrification in one bioreactor is a possibility.

How one can obtain full control over the daily nitrate uptake rate using carbohydrates?

The most simple and easily applied method for removing nitrate and on which I have done some research, suitable for having control over the nitrate level , is the SPC method, sulphur packed columns. A mix of oystershell gravel and elemental sulphur beats putted in a filter bag and hung freely in the water column. Very cheap, very effective and very low maintenance. http://www.baharini.eu/baharini/doku...lommen_systeem

[Dan_P]

Originally Posted by Tripod1404

Yeah that is the question. I actually accidentally realized this. I was trying to raise nitrates by dosing amino acids, but more I dosed more it reduced ( I actually dropped to undetectable NO3 levels on salifert kit). Than I decided something was not right, I stopped dosing aminos and nitrate slowly started to increase.

Interesting observation!

[Dan_P]

Originally Posted by Tripod1404

I use a over sized skimmer, so I am very cautious about how much skimming is going on. Aside from that it causes the skimmer to foam like crazy and overflow, if I dont turn it off.

How many ppm of amino acids are you adding? Which ones seem to cause the most foam?

[Tripod1404]

Originally Posted by Dan_P

How many ppm of amino acids are you adding? Which ones seem to cause the most foam?

I dont prepare my amino acid stock solutions. I use commercial amino acid supplements like; acropower, KZ LPS aminos acids and red sea reef energy B.

I generally dose 10-15ml of one of these products to ~120G tank daily. If that days nitrate tests gave less than ~0.5ppm nitrate, I dose half the amount. Among these acropower cause the most foaming. Even after 1 hour of turning the skimmer off, it still foams if what is dosed is acropower. I think this is because acropower is the most concentrated product among these. Manufacturer Recommended dosing amount of AP is ~1/10 of the other brands(but it still dose 10-15ml).

[Tripod1404]

Originally Posted by Belgian Anthias

How difficult and expensive one can make a simple task as removing non toxic nitrate?

Do we want to remove the nitrogen or assimilate and recycle it?
Depending of the daily quantity nitrate to remove.
Do we want to keep the present nitrification capacity or shift the ammonium reduction towards organic carbon availability and bacterial growth?
Labor-intensive or normal maintenance practice?
Are all side effects known and how they are prevented?

Removing nitrate? Just controlling the nitrate level? Or do we want to have full control over the daily nitrate removal rate and close the nitrogen cycle?

For removing nitrate from the system we have to reduce it to nitrogen gas.
Controlling the nitrate level is possible both ways, denitrification or assimilation, depending of how much nitrate has to be reduced on a daily base. ( preventing bacterial slime, high C:N ratio's)
For full control over the daily nitrate removal rate one needs a bioreactor and have control over the HRT, the flow rate.
Simultane nitrification and mixotropic denitrification in one bioreactor is a possibility.

How one can obtain full control over the daily nitrate uptake rate using carbohydrates?

The most simple and easily applied method for removing nitrate and on which I have done some research, suitable for having control over the nitrate level , is the SPC method, sulphur packed columns. A mix of oystershell gravel and elemental sulphur beats putted in a filter bag and hung freely in the water column. Very cheap, very effective and very low maintenance. http://www.baharini.eu/baharini/doku...lommen_systeem

Well ,removing nitrate from the system does not require it to be reduced to nitrogen gas. Whole bacteria can be skimmed out of the tank, together with nitrogen containing bio-molecules, about 3-5% of a cell is nitrogen anyways. I am skeptical about carbon dosing increasing nitrate reduction into nitrogen gas. That process is severely inhibited by oxygen. So if we assume it only happens at regions that oxygen cannot diffuse, we should also assume larger organic carbon sources wont diffuse to these bacteria as well. I have read interesting theories about carbon dosing increasing the oxygen consumption by bacteria, forcing them (especially the ones living deeper within the bacterial mat) to become facultatively anaerobic for some period. But I doubt this state would last long enough to have signification reduction into N2 gas.

In my opinion carbon dosing causes bacteria population to increase. Building more bacteria requires more nitrogen, so ammonia/nitrite/nitrate is taken up by growing bacteria. Some of these bacteria gets waterborne and skimmed out by the skimmer, effectively exporting nitrogen out of the system.

I get what you mean by saying this process makes nitrogen export dependent organic carbon availability. But the alternative you described is not that different. The only difference is you make the process dependent on elemental sulfur availability. By giving them sulfur to reduce, you allow them to chemotrophicly fix carbon dioxide. So in some ways that is indirect carbon dosing . Similar to growing algae and allowing them to fix carbon phototrophicly and absorb nitrogen for growth.

In the end an aquarium is a closed system. As much as we want it to be, it is not an ecosystem, it is not self sufficient. In a ecosystem biomass is composed of autotrophs (mainly plants and algae) >> simple heterotrophs (bacteria, fungi and other microorganisms) >> complex heterotrophs (animals). In an aquarium, the biomass is almost flipped, we have complex heterotrophs > simple heterotrophs >> autotrophs. Such a system cant be functioning without external input.

[bertoni]

I think you meant, "an aquarium is not a closed system"?

I agree that nitrogen gas is just one path out of the system for nitrogen. Skimming bacteria or bacterial output both could be important. I have read differing opinions on which is more likely in the skimmate.

[Tripod1404]

Originally Posted by bertoni

I think you meant, "an aquarium is not a closed system"?

I agree that nitrogen gas is just one path out of the system for nitrogen. Skimming bacteria or bacterial output both could be important. I have read differing opinions on which is more likely in the skimmate.

Yeah not a closed system