The Bacterial “Rip Clean” Method

[ReefGeezer]

While I have no experience with what now seems to be subject of the thread, I do feed organic carbon in several ways. I dose some vinegar (acetate), phytoplankton, and sometimes Reef Energy AB+. The latter is very high in carbohydrates, but I'm not sure they are close to the sugars being spoken of in this thread.

I've found that it is easy to over do the Reef Energy AB+. I think that while small amounts might encourage useful bacteria, it might also be true that too much feeds pest bacteria like Cyano. Could that also be true with Reef Actif?

 

[sixty_reefer]

I think what I'm going to be doing for my tank will be quarter doses nightly for the first week then move to half doses for the second week nightly. I will reassess from there and see if anything is happening.
If 3g is a spoon, I need 75% as full dose divided by 4.
2.25g / 4 = .56g... Sound right? Sound too aggressive to start?

Since somebody mentioned the skimmer and do have a question. Should it be running while we dose? Should I shut it off for an hour after dose or the whole night after Dose or don't bother shutting it off at all?

I think @Subsea is using half dose with no issues, he may want to chime in here.
Leave everything else as normal including protein skimmer.

 

[Dburr1014]

I think @Subsea is using half dose with no issues, he may want to chime in here.
Leave everything else as normal including protein skimmer.

To clarify a point about me going to 1/2 dose protocoal.
Actually, I am dosing at 50% of maximum dose. This 75G display has many diverse filter feeders at 25 years mature.

Also, with respect to dosing three times a week, I dose every 2 days.

@Subsea is this your starting dose?

 

[sixty_reefer]

While I have no experience with what now seems to be subject of the thread, I do feed organic carbon in several ways. I dose some vinegar (acetate), phytoplankton, and sometimes Reef Energy AB+. The latter is very high in carbohydrates, but I'm not sure they are close to the sugars being spoken of in this thread.

I've found that it is easy to over do the Reef Energy AB+. I think that while small amounts might encourage useful bacteria, it might also be true that too much feeds pest bacteria like Cyano. Could that also be true with Reef Actif?

From what o read from the manufacturer claims and from the paper in the opening post the biggest difference is the number of C atoms on the carbohydrate. We’re more complex chains will make it harder to be used by nuisance organisms.
Actif is supposed to be a complex carbohydrate and that may be the biggest difference imo

quote from the paper.

molasses are rapidly soluble in water and can be quickly absorbed by heterotrophic bacteria 3. In terms of chemical structure, molasses are classified as a simple carbohydrate containing six C atoms (monosaccharides), while sucrose (treatment D) is a combination of two monosaccharides that contain 12 C atoms (sucrose). Tapioca is classified as a complex carbohydrate (60,000 C atoms) and is more slowly digested by bacteria than molasses.

 

[Randy Holmes-Farley]

From what o read from the manufacturer claims and from the paper in the opening post the biggest difference is the number of C atoms on the carbohydrate. We’re more complex chains will make it harder to be used by nuisance organisms.

quote from the paper.

molasses are rapidly soluble in water and can be quickly absorbed by heterotrophic bacteria 3. In terms of chemical structure, molasses are classified as a simple carbohydrate containing six C atoms (monosaccharides), while sucrose (treatment D) is a combination of two monosaccharides that contain 12 C atoms (sucrose). Tapioca is classified as a complex carbohydrate (60,000 C atoms) and is more slowly digested by bacteria than molasse

I have wings and can fly.

Don't believe everything you read.

Molasses is mostly sucrose. See table 1 in the paper below.

https://www.journalofdairyscience.org/article/S0022-0302(20)30307-6/pdf

 

[Randy Holmes-Farley]

While I have no experience with what now seems to be subject of the thread, I do feed organic carbon in several ways. I dose some vinegar (acetate), phytoplankton, and sometimes Reef Energy AB+. The latter is very high in carbohydrates, but I'm not sure they are close to the sugars being spoken of in this thread.

The term carbohydrate covers millions of different structures, and is even abused by some reefers to include acetic acid.

 

[sixty_reefer]

I have wings and can fly.

Don't believe everything you read.

Molasses is mostly sucrose. See table 1 in the paper below.

https://www.journalofdairyscience.org/article/S0022-0302(20)30307-6/pdf

That’s correct mostly sucrose with a little fructose and glucose. I looked into brown sugar after reading Beuchat thread on dinoflagellates to try and understand why brown sugar seemed to be more effective in comparison to only use only ethanol and acetic acid.
Although sucrose, fructose and glucose are still considered simple carbohydrates they still have longer chains in comparison to ethanol and acetic acid (not carbohydrates) the longer chains claimed by TM may have some grounds into being more difficult to be metabolised by some nuisances. Maybe something that we as reefers should look into as we all know that wile dosing acetic acid and ethanol Cyanobacteria sometimes seems to be able to use it.

 

[ReefGeezer]

The term carbohydrate covers millions of different structures, and is even abused by some reefers to include acetic acid.

I simply imagine that reefers abuse scientific terms or that there is gambling in River City. Yes, I'm that old!

 

[taricha]

I was actually surprised that the 3 sources of carbon in the paper from the OP were as similar as they were in terms of the measurable levels of Ammonia/NO2/NO3.
That no statistically significant differences were found between sucrose, molasses, and tapioca additions, might suggest that bacteria can adjust to a wide range of carbon sources - or that those sources are not as different as they would appear. If you have organisms that can chop up starch, then it's not much different from a simple sugar I guess.

 

[beesnreefs]

Actif just arrived and I plan to give this a test run. My hope is this helps me eliminate the cyano I've got matting up on my sand these past few weeks.

I'll be started with 1 level scoop (to treat 132 gallons) every 2 days to mimic what @Subsea is doing (and match @sixty_reefer recommendations). My tank is a 225 gallon system so this is roughly half the maximum dose. First dose will go in tonight after lights out.

Before pictures of the tank just taken:

Full tank just before ramp down begins.

Some of the GHA that has started to grow in the past 2 weeks

Front glass about 2 days after last getting scraped.

This is the area I will not siphon during this experiment. I'll leave it be to see what, if any, changes are detected.

 

[sixty_reefer]

Actif just arrived and I plan to give this a test run. My hope is this helps me eliminate the cyano I've got matting up on my sand these past few weeks.

I'll be started with 1 level scoop (to treat 132 gallons) every 2 days to mimic what @Subsea is doing (and match @sixty_reefer recommendations). My tank is a 225 gallon system so this is roughly half the maximum dose. First dose will go in tonight after lights out.

Before pictures of the tank just taken:

Full tank just before ramp down begins.

Some of the GHA that has started to grow in the past 2 weeks

Front glass about 2 days after last getting scraped.

This is the area I will not siphon during this experiment. I'll leave it be to see what, if any, changes are detected.

Could you post a pic every few days of the area that you not touching the Cyanobacteria? Also are your residual nitrates and phosphate detectable?

 

[beesnreefs]

Could you post a pic every few days of the area that you not touching the Cyanobacteria? Also are your residual nitrates and phosphate detectable?

Yes I will post pictures as frequently as I can. I’ll be out of town the next two days but will post again on Thursday

Nitrates and phosphates are consistently detectable in this tank. Nitrates hold fairly steady at 3-5 ppm and phosphates are consistently between 0.05 and 0.1

 

[littlefishy]

If the normal routine is removal of cyano and scraping of glass, shouldn't that and all routines be maintained so variables are eliminated? I have ots in my filterless 10g w cyano as the main algae and have considred this experiment instead of a combo of PaulB's canister blasting of the rocks and Brandon's sandbed removal and cleanse.

 

[beesnreefs]

If the normal routine is removal of cyano and scraping of glass, shouldn't that and all routines be maintained so variables are eliminated? I have ots in my filterless 10g w cyano as the main algae and have considred this experiment instead of a combo of PaulB's canister blasting of the rocks and Brandon's sandbed removal and cleanse.

FWIW, I intend to continue with my normal routine. I'll conduct a water change this weekend like I always do (roughly 8%), scrape the glass, and siphon cyano except the small section I noted in my post above.

 

[Subsea]

I think @Subsea is using half dose with no issues, he may want to chime in here.
Leave everything else as normal including protein skimmer.

I am starting at 75% of recommended maximum dosage on healthy 75G 25 year mature tank. On 120G tank , I am going full dose of 3 grams. and dosing every 2 days. Administered the third treatment 6 hours ago.

 

[beesnreefs]

I am starting at 75% of recommended maximum dosage of healthy 75G 25 year mature tank. On 120G tank , I am going full dose of 3 grams. and dosing every 2 days. Administered the third treatment 6 hours ago.

Thanks for clarifying, @Subsea.

Did you start at these dosages right away? Or work up to them?

 

[taricha]

For those interested in the sorts of biopolymers involved see this from Hans-Werner for some discussion.

To be totally correct there is some overlap between Tropic Marin Reef Actif and NP-Bacto-Pellets, but there are also big differences although we tried to address diverse and similar organisms with both products and have tried to include some important similarities.

NP-Bacto-Pellets contain no algal biopolymers, but we included other biopolymers than the bacterial biopolymer PHA/PHB into the pellets to maybe feed heterotroph microbes that form DHA, an important marine long-chain polyunsaturated fatty acid which is meant to be necessary for the normal development of some marine organisms, fish fry amongst others.

I am sorry but I have to say alginate is not a protein. In fact alginate is one of the marine biopolymers included in Reef Actif but only to a small proportion. Alginate in not a protein and it does not contain nitrogen and I think it is not an ingredient of normal fish feeds. Alginate is a polysaccharide with charged groups that can bind calcium and other metals and give it quite unique properties.

(This is not Subsea's goal, but...)
If what somebody wanted to do was to reduce the availability of N forms (ammonia/NO2/3) to nuisance growth, then "fancy" carbon sources are not necessarily required.
You can slam systems with high amounts of vinegar or vodka and the nuisance growth will fade - figuratively and literally as the pigment composition shows N-limitation.

Here's where I did exactly such an exercise a couple of years ago.

Why Do Large Carbon Doses Suppress Cyano (in my system)?
A big part of this method is about using bacteria to digest the waste that fuels nuisance growth, and the other big part is about using large carbon dose to tweak bacterial behavior.
In the spirit of taking things apart, I wanted to take a closer look at the large carbon dose part, and why large doses of organic carbon suppress cyano growth in my system. It appears to disrupt cyano's ability to exploit accumulations of waste.

Usually in my tank, algae dying (from vibrant) results in debris material that supports cyano growth, but if I do a large daily carbon dose (while keeping vibrant), the cyano recedes despite the debris source still being produced.
So I turned cyano off/on/off to investigate.

(left: while dosing carbon no cyano on dying algae, middle: no carbon dosing and lots of red cyano, right: heavy carbon dosing and re-disappearance of cyano. The carbon dosing here is 2/3 of the carbon dose recommended in the method.)
Bulk water parameters were not dramatically changed. PO4 stayed around ~0.10 and NO3 tests zero...if you don't look too closely. More on that in a bit.

So why is the cyano receding?
In the sump there's an interesting stringy cyano colony that goes through color shifts when I dose large amounts of carbon. It goes from deep emerald green -> light green -> yellow, and when carbon dosing stops, it reverses.
This sort of color transition in cyano has been known for decades to be associated with N depletion.
"...the alga [cyanobacteria] lost its characteristic blue-green color and took on the light brown to golden color of a diatom. Experimentation showed that it was not the wavelength of light, but rather the depletion of nitrogen from the medium, and that alone, which was responsible for the color shift. In this and further work ..., it was determined that the color shift was due to a selective disappearance of chlorophyll and phycocyanin, thereby unmasking the carotenoids; that a yellowed culture could be made to return to green, sometimes within 24 hours, upon the addition of minute amounts of nitrogen..."
paper link
Other research in the 60 years since, has fleshed out how in nutrient deprivation, the cells become photobleached, and reduce their light-harvesting pigments while keeping more photoprotective carotenoids.

So I ran some spectrum test on samples from the cyano during increased carbon dosing, and I found the exact pigment shifts described above that are associated with N depletion.

On the left is the colony of cyano, on the right are the spectral measurements from the sample. Black is the actual measured spectra, grey dotted line is a fit made from the pigments listed in the graph header. Different colored lines represent different pigments that add up to the gray dotted line.
Top: emerald green cyano - 10/21 - note the strong phycocyanin (blue line).
Middle: light-green - 10/25 - Phycocyanin almost collapsed and Chlorophyll A is slightly reduced compared to carotenoids.
Bottom: yellow - 10/28 - total collapse of phycocyanin, and Chlorophyll A reduced compared to carotenoids which become dominant.

Additionally, the pigments not only shifted, the overall pigment density of these samples decreased during this process. Below compares absorption from light-harvesting pigments, to the scattering by the material after bleached with a couple of drops of sodium hypochlorite.

Solid lines are raw absorbance (normalized to 1) and the dotted lines represent what scattering remained when the sample was bleached.
The 10/21 - emerald green sample - blue line had over half 58% of its absorbance from actual pigments.
10/25 - light-green sample - working pigments had dropped to 40%
10/28 - yellow sample - pigment absorbance was only 13% of sample.

This indicates that by the end there was severe light-harvesting pigment loss, major photobleaching of the sample. It suggests that a lot of the sample was dead at this point (the strands detached from the pipe 4 days later) , All this data points to nutrient (Nitrogen) deprivation.

To be clear, I'm not saying this is desirable. Or sustainable in a reef tank.
Just saying it does happen.

More sophisticated carbon sources may be desirable in a number of ways.

I'd like a carbon source that specifically drove skimmable bacterial growth in the water - if I were daydreaming my organic carbon ideal.

 

[Subsea]

Thanks for clarifying, @Subsea.

Did you start at these dosages right away? Or work up to them?

I started with first dose.

I choose a reduced dose rate for 75G tank, because it is in good shape except for persistent cynobacteria that seems to be ChemiClean resistant.

 

[Subsea]

For those interested in the sorts of biopolymers involved see this from Hans-Werner for some discussion.




(This is not Subsea's goal, but...)
If what somebody wanted to do was to reduce the availability of N forms (ammonia/NO2/3) to nuisance growth, then "fancy" carbon sources are not necessarily required.
You can slam systems with high amounts of vinegar or vodka and the nuisance growth will fade - figuratively and literally as the pigment composition shows N-limitation.

Here's where I did exactly such an exercise a couple of years ago.


To be clear, I'm not saying this is desirable. Or sustainable in a reef tank.
Just saying it does happen.

More sophisticated carbon sources may be desirable in a number of ways.

I'd like a carbon source that specifically drove skimmable bacterial growth in the water - if I were daydreaming my organic carbon ideal.

Are you in academia or industry?

PS: My overall goal is to understand why and as I learn more, I realize how little I know. Thank you for helping to tip the scales.

 

[Subsea]

If it matters so much then why is he unable to say what the best source is? Does he say what the "best" source would be?

Not sure how to best answer the tone of your post.

With respect to perfect solutions, they don’t exist. There are trade offs with pros & cons.

With respect to biofloc in intensive shrimp farms, you should ask the doctor at his consulting business.

Dr. Tzachi Samocha​

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