Carbon dosing no bacteria film but nitrates dropping

[Miami Reef]

I am carbon dosing a mixture of 75% vinegar and 25% vodka. I used 25mL a day in my 260 gallon. Nitrates dropped to 0ppm. I dosed 4ppm and 2 days later nitrates dropped back to 0ppm.

I dosed to 2ppm and reduced dosage to 10mL per day.

I’m trying to grow bacteria on my rocks, glass, sump because I eventually want to try a starfish but I’m not seeing any bacterial growth.

I feel like lowering my dose will just delay the bacterial process, but I cannot keep at my old dose since nitrates are dropping too fast.

Anyone with carbon dosing experience have an idea to get more bacteria on rocks/sand?

I use a mix of dry and ocean rock (KP) and it’s been running for about 6 months.

 

[kdtorgy]

My understanding (and I could be wrong) is the bacteria produced by carbon dosing are pelagic and live in the water column and don’t colonize surfaces. I have high nitrates and phosphates. Tried carbon dosing and killed my yellow tang. Trying PNS ProBio which are supposed to colonize surfaces and reduce nutrients. Just started a week ago so a little too early to tell. I know people have also used microbacter 7 from brightwell.

PNS ProBio™

By removing ammonia, nitrite and nitrate, consuming organic waste products, providing a nutritious food source for filter-feeders and acting as a powerful probiotic, PNS ProBio™ is a highly beneficial addition to your aquarium's microbial community. This product is especially useful for feeding...

www.algaebarn.com

Brightwell Aquatics - Microbacter7

 

[taricha]

I do a similar ratio sometimes 8:1 vinegar:vodka and it produces visible bacterial growth on surfaces at 20mL in 60 gal volume, whether I have detectable nitrates or not.
The nutritional content of what's grown may be low if there's virtually no N available during the formation. No idea.

Just because there's no visible growth doesn't mean no growth. bacterial films being colorless are not obvious.

 

[Dan_P]

I am carbon dosing a mixture of 75% vinegar and 25% vodka. I used 25mL a day in my 260 gallon. Nitrates dropped to 0ppm. I dosed 4ppm and 2 days later nitrates dropped back to 0ppm.

I dosed to 2ppm and reduced dosage to 10mL per day.

I’m trying to grow bacteria on my rocks, glass, sump because I eventually want to try a starfish but I’m not seeing any bacterial growth.

I feel like lowering my dose will just delay the bacterial process, but I cannot keep at my old dose since nitrates are dropping too fast.

Anyone with carbon dosing experience have an idea to get more bacteria on rocks/sand?

I use a mix of dry and ocean rock (KP) and it’s been running for about 6 months.

What kind of sea star are you considering and what is the connection with a bacteria film?

 

[livinlifeinBKK]

Any biofilm you successfully grow in a tank is going to vary GREATLY compared to the natural biofilm diet of a starfish that eats biofilm in the wild

 

[livinlifeinBKK]

I had a similar idea but after researching marine biofilms, I abandoned the idea. Not only does the water chemistry and nutrient content affect the biofilm that begins to form, but simple environmental factors can change the composition dramatically...not to mention that nobody knows exactly what it is in the biofilm that they actually need for their nutrition.

 

[Miami Reef]

What kind of sea star are you considering and what is the connection with a bacteria film?

Linkia Starfish.

 

[Randy Holmes-Farley]

FWIW, I never saw benthic bacteria when dosing vodka or vinegar, even at very high doses, even to the point where the water became hazy with bacteria, but I had lots of rocks in my refugia for them to spread out.

 

[ISpeakForTheSeas]

Getting the bacteria propagating is one thing - getting them to form a biofilm is another, as they typically form biofilms in response to some sort of environmental stressor ("such as UV radiation, desiccation, limited nutrients, extreme pH, extreme temperature, high salt concentrations, high pressure, and antimicrobial agents.")*, and what stressor causes each bacteria species to form a biofilm may vary. There are also a number of other factors that influence the biofilm formation and composition as well (including what microbe species are in the aquarium, flow, nutrients, substrate composition and porosity, lighting, oxygen levels, etc.).

As Taricha mentioned, there may be substantial biofilm formation that's just not visible without intensive (and quite possibly microscopic) inspection. Importantly, however, as was also mentioned, we don't know what is nutritionally needed in these biofilms for these starfish (and biofilms can be very simple or very complex in nature, with most that I've seen actually analyzed being rather complex), so even if you get successful biofilm propagation in your aquarium, there is no guarantee that it will contain the right amounts of the right "ingredient or ingredients" (bacteria, diatoms, fungus, etc.) in it for starfish health.

This is not to try and discourage you, it's just to let you know that this may not be a simple task.

*This article is where I pulled the quote from above, and it discusses a few biofilm forming environmental stressors in some depth: https://www.frontiersin.org/articles/10.3389/fmicb.2020.00928/full#h5

 

[ISpeakForTheSeas]

*This article is where I pulled the quote from above, and it discusses a few biofilm forming environmental stressors in some depth: https://www.frontiersin.org/articles/10.3389/fmicb.2020.00928/full#h5

Sorry, slight correction to my statement here, it discusses a few biofilm forming environmental influences, not stressors, in some depth. It discusses the stressors briefly (as quoted above) and has links to the articles that discuss the environmental stressors in more depth. Apologies for my mis-typing there.

 

[Dan_P]

Getting the bacteria propagating is one thing - getting them to form a biofilm is another, as they typically form biofilms in response to some sort of environmental stressor ("such as UV radiation, desiccation, limited nutrients, extreme pH, extreme temperature, high salt concentrations, high pressure, and antimicrobial agents.")*, and what stressor causes each bacteria species to form a biofilm may vary. There are also a number of other factors that influence the biofilm formation and composition as well (including what microbe species are in the aquarium, flow, nutrients, substrate composition and porosity, lighting, oxygen levels, etc.).

As Taricha mentioned, there may be substantial biofilm formation that's just not visible without intensive (and quite possibly microscopic) inspection. Importantly, however, as was also mentioned, we don't know what is nutritionally needed in these biofilms for these starfish (and biofilms can be very simple or very complex in nature, with most that I've seen actually analyzed being rather complex), so even if you get successful biofilm propagation in your aquarium, there is no guarantee that it will contain the right amounts of the right "ingredient or ingredients" (bacteria, diatoms, fungus, etc.) in it for starfish health.

This is not to try and discourage you, it's just to let you know that this may not be a simple task.

*This article is where I pulled the quote from above, and it discusses a few biofilm forming environmental stressors in some depth: https://www.frontiersin.org/articles/10.3389/fmicb.2020.00928/full#h5

I read the paper. I think the biofilm formation quote should be read as
”biofilm formation can be formed in response to stress” because biofilms are a normal way of life for benthic bacteria. Biofilms have so many advantages for bacteria. It would be next to impossible to stop their formation.

Another thing about bacteria biofilms is that they are not pure. Many bacteria species live together in a.biofilm and photosynthetic microorganisms are associated with it as well. And there are all sorts of higher life forms harvesting these microorganisms. When a sea star is “consuming a biofilm”, it is likely slurping up many species of microorganisms.

As for what bacteria consume acetic acid or ethanol in an aquarium, pelagic, benthic or both, has anyone actually studied this? I ask because I can grow white film on surfaces in my system and some of those films contain large, clear, segmented (visible cell walls) filaments (I never attempted to ID. I was like “oh wow” and moved away from that rabbit hole).

 

[taricha]

I had a similar idea but after researching marine biofilms, I abandoned the idea. Not only does the water chemistry and nutrient content affect the biofilm that begins to form, but simple environmental factors can change the composition dramatically...not to mention that nobody knows exactly what it is in the biofilm that they actually need for their nutrition.

All true.
But in the spirit of throwing out some ideas just for discussion...

I'd probably try a mix like this:
glucose, acetate, glutamine, and live phytoplankton.
Reasoning: Glucose is the majority organic carbon released by a lot of macroalgae.
And acetate is a widespread and widely used carbon source on reefs, by everything from bacteria to the coral animal. Also convenient.
Glutamine is an amino acid with high N content. I have no reason to prefer it over other aminos except it has a lot of N. Some organisms will consume it whole, others will break it down and re-release some N as ammonia. So it can also be thought of as a slow release ammonia source. In my system, adding glutamine, I can accumulate NO3 (undetectable otherwise). Someone who's thought more about amino acid behavior in marine systems will probably have a different answer for a good amino acid choice for substrate-dweller health.
And then phytoplankton (T-iso for high nutrition, or nanno for convenience of culturing) because all other things listed are only foods to organisms that consume nutrition dissolved in the water. Many organisms are filter feeders.

I would guess something like that would create robust live layers across every square cm of rock. Though what's grown may not be desirable for a general reef tank. And no guarantees that what's grown would even be good for grazers of substrate.

I have no thoughts on ratios of those except I'd manage the carbon to amino acid ratio in a way to keep a small bit of detectable NO3 endpoint in the water.

 

[Dan_P]

All true.
But in the spirit of throwing out some ideas just for discussion...

I'd probably try a mix like this:
glucose, acetate, glutamine, and live phytoplankton.
Reasoning: Glucose is the majority organic carbon released by a lot of macroalgae.
And acetate is a widespread and widely used carbon source on reefs, by everything from bacteria to the coral animal. Also convenient.
Glutamine is an amino acid with high N content. I have no reason to prefer it over other aminos except it has a lot of N. Some organisms will consume it whole, others will break it down and re-release some N as ammonia. So it can also be thought of as a slow release ammonia source. In my system, adding glutamine, I can accumulate NO3 (undetectable otherwise). Someone who's thought more about amino acid behavior in marine systems will probably have a different answer for a good amino acid choice for substrate-dweller health.
And then phytoplankton (T-iso for high nutrition, or nanno for convenience of culturing) because all other things listed are only foods to organisms that consume nutrition dissolved in the water. Many organisms are filter feeders.

I would guess something like that would create robust live layers across every square cm of rock. Though what's grown may not be desirable for a general reef tank. And no guarantees that what's grown would even be good for grazers of substrate.

I have no thoughts on ratios of those except I'd manage the carbon to amino acid ratio in a way to keep a small bit of detectable NO3 endpoint in the water.

If I were a bacterium, I would eat @taricha home style tangy medium of glucose and glutamine in vinegar!

Managing the recipe by nitrate in the water sounds reasonable and simple. I think you can calculate the ratio by how much carbon goes to energy and how much to biomass.

 

[livinlifeinBKK]

Btw, when I said I abandoned the idea, I meant the idea of growing a suitable biofilm using only additives (I do still intend to try to come up with an alternative food source)

 

[ISpeakForTheSeas]

I read the paper. I think the biofilm formation quote should be read as
”biofilm formation can be formed in response to stress” because biofilms are a normal way of life for benthic bacteria. Biofilms have so many advantages for bacteria. It would be next to impossible to stop their formation.

Another thing about bacteria biofilms is that they are not pure. Many bacteria species live together in a.biofilm and photosynthetic microorganisms are associated with it as well. And there are all sorts of higher life forms harvesting these microorganisms. When a sea star is “consuming a biofilm”, it is likely slurping up many species of microorganisms.

As for what bacteria consume acetic acid or ethanol in an aquarium, pelagic, benthic or both, has anyone actually studied this? I ask because I can grow white film on surfaces in my system and some of those films contain large, clear, segmented (visible cell walls) filaments (I never attempted to ID. I was like “oh wow” and moved away from that rabbit hole).

That's fair - many benthic bacteria would likely only go planktonic as a result of extreme stress. However, institutions also culture many of these benthic, biofilm-forming species in their planktonic forms, so it's possible (though, as hinted at above, I agree it's unlikely) that even a number of benthic bacteria species need the proper environment/conditions to be encouraged into biofilm formation (advantageous though it generally may be).

I touched on the point of them not being pure above (they can be pure, but, to my knowledge, we have to culture them that way intentionally for them to be so - their usual compositions are extremely diverse) with my statement here:
"... we don't know what is nutritionally needed in these biofilms for these starfish (and biofilms can be very simple or very complex in nature, with most that I've seen actually analyzed being rather complex), so even if you get successful biofilm propagation in your aquarium, there is no guarantee that it will contain the right amounts of the right "ingredient or ingredients" (bacteria, diatoms, fungus, etc.) in it for starfish health."

I have no idea if someone has attempted to study which bacteria consume our carbon dosing additives - that's an interesting thought, and I would love to see if one type of carbon dosing is preferred by different types of bacteria.

All true.
But in the spirit of throwing out some ideas just for discussion...

I'd probably try a mix like this:
glucose, acetate, glutamine, and live phytoplankton.
Reasoning: Glucose is the majority organic carbon released by a lot of macroalgae.
And acetate is a widespread and widely used carbon source on reefs, by everything from bacteria to the coral animal. Also convenient.
Glutamine is an amino acid with high N content. I have no reason to prefer it over other aminos except it has a lot of N. Some organisms will consume it whole, others will break it down and re-release some N as ammonia. So it can also be thought of as a slow release ammonia source. In my system, adding glutamine, I can accumulate NO3 (undetectable otherwise). Someone who's thought more about amino acid behavior in marine systems will probably have a different answer for a good amino acid choice for substrate-dweller health.
And then phytoplankton (T-iso for high nutrition, or nanno for convenience of culturing) because all other things listed are only foods to organisms that consume nutrition dissolved in the water. Many organisms are filter feeders.

I would guess something like that would create robust live layers across every square cm of rock. Though what's grown may not be desirable for a general reef tank. And no guarantees that what's grown would even be good for grazers of substrate.

I have no thoughts on ratios of those except I'd manage the carbon to amino acid ratio in a way to keep a small bit of detectable NO3 endpoint in the water.

Sounds well thought out. T-iso and Nanno is likely a good place to start. I would add Chaetoceros gracilis too, but I don't know if it would help or not.

 

[taricha]

As for what bacteria consume acetic acid or ethanol in an aquarium, pelagic, benthic or both, has anyone actually studied this? I ask because I can grow white film on surfaces in my system and some of those films contain large, clear, segmented (visible cell walls) filaments (I never attempted to ID. I was like “oh wow” and moved away from that rabbit hole).

That's entirely unstudied to my knowledge. Mostly we just calculate equivalent carbon and dose them interchangeably, even though if you do this, the visible growth from different carbon sources are naked-eye-obviously different.

 

[sixty_reefer]

I am carbon dosing a mixture of 75% vinegar and 25% vodka. I used 25mL a day in my 260 gallon. Nitrates dropped to 0ppm. I dosed 4ppm and 2 days later nitrates dropped back to 0ppm.

I dosed to 2ppm and reduced dosage to 10mL per day.

I’m trying to grow bacteria on my rocks, glass, sump because I eventually want to try a starfish but I’m not seeing any bacterial growth.

I feel like lowering my dose will just delay the bacterial process, but I cannot keep at my old dose since nitrates are dropping too fast.

Anyone with carbon dosing experience have an idea to get more bacteria on rocks/sand?

I use a mix of dry and ocean rock (KP) and it’s been running for about 6 months.

Could you not just add more fish to your system, feed more and then balance that load with carbon dosing?