Analyzing a Bacterial Method for Dinoflagellates (and cyano?)

Victoria M

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Thank you so much for sharing this with us! I get so excited when I see you post @taricha . Like fan girl excited. Please carry on gentleman. Steps back quietly...


I presume that Dr. Tims studies to prove his theses on bacteria ran on very similiarly lines with tests tubes, grunge and his specifically isolated bacteria ...
I would love to read that paper.
 

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Best thing I know of along these lines is still Feldman article.
Bacterial Counts in Reef Aquarium Water: Baseline Values and Modulation by Carbon Dosing, Protein Skimming, and Granular Activated Carbon Filtration
To do better than this, I think you'd have to know something about which bacteria you are seeing are good/bad and I have no idea if that's doable.
its not.

Not in a way that preserves the relative numbers or even lets you quantify them in any way. It would be a present/absent qualifier, and then only if you could get 100% of the bacteria present to grow on your media.

Pro-tip: you can’t.

The only other way to do it would be environmental whole genome screening but no one is paying the $10k/sample data acquisition and they $80/hour x 40 hours for analysis and assembly.

I’ve done it when pulling an infected pancreas from a patient to look at pathogenic bacterial colonization and relative numbers, but that was part of a type 1 diabetes pancreatitis study. I don’t think anyone is paying that for aquarium ecology.
 

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I’d be very interested in seeing a comparison of the bacterial counts, of all different types of bacteria, found in the water column of both “healthy” reef tanks, as well as those with an over abundance of problematic Dinoflagellates. The bacteria found on surfaces, and in the substrate, as well.

Oh, and how the numbers compare before and after running the regimen to “the T”
Unfortunately, impossible.

Counts rely on being able to grow the bacteria into a visual colony. Currently, we can only guess, but our present ability to grow environmental bacteria is limited to around 9-30% of strains present.
 

JAMSOURY

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Just a quick update, 2 weeks after doing the method 3 times, no trace of dinos yet! Keeping close watch and raising my nutrients after all the bacteria took out all my nitrates. Phosphates at 0.04. All sps are starting to get their color probably after all the irritation it has gone through the process
 

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Well, this has been a very interesting read... although I did skip the sections where the PhD's battled it out over Henry's Law, etc...

Thanks @taricha for putting the effort into this and all your other dino experiments! It is greatly appreciated. We desperately need a solution to the amphidinium dino problem!!

I battled these dinos for almost the past year by elevating nitrates, elevating phosphates, adding MicroBacter7 sporadically, dosing silica, adding pods, etc... I finally pushed the dinos back to just a fine dusting on my sand bed in a few spots. Then, the green cyano covered my tank! Expected with such a crazy fluctuation in nutrients. I feared dropping my No3/Po4 levels too quickly to get the cyano under control would bring back the dinos. So, I reduced nutrients slowly to around 5ppm No3 and .06ppm Po4... and sucked out cyano weekly after it would coat my rocks again.

Well, everything looked good for awhile... but the dinos are increasing again on my sand bed! Well, the dinos are taking hold in areas of my sand where I had dumped sand back into my tank after "cleaning" it. During my siphoning of cyano out, I would usually get about 1-2 cups of sand as well... so I rinsed that sand with fresh water to lyse the dinos (since I would also siphon out areas with them in it as well)... and also treated that sand with some H2O2 to further kill dinos and cyano... then let it completely dry and then returned to the tank. Well, those areas are where the dinos are coming back! I think by cleaning the sand, it created a very sterile environment where the dinos had no competition; therefore, they are starting to flourish there. I wonder if after cleaning this sand, treating it by soaking it in a nitrifying bacteria solution (Microbacter7 + saltwater, etc...) and then returning it to the tank would keep the dinos from getting a foothold in the cleaned sand? Hmmm...

But, I think I'm going to give the Elegant Coral procedure a try. After so many months of battling with other treatments... I'm tired. A final decisive blow would be refreshing...
 

Victoria M

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Well, this has been a very interesting read... although I did skip the sections where the PhD's battled it out over Henry's Law, etc...

Thanks @taricha for putting the effort into this and all your other dino experiments! It is greatly appreciated. We desperately need a solution to the amphidinium dino problem!!

I battled these dinos for almost the past year by elevating nitrates, elevating phosphates, adding MicroBacter7 sporadically, dosing silica, adding pods, etc... I finally pushed the dinos back to just a fine dusting on my sand bed in a few spots. Then, the green cyano covered my tank! Expected with such a crazy fluctuation in nutrients. I feared dropping my No3/Po4 levels too quickly to get the cyano under control would bring back the dinos. So, I reduced nutrients slowly to around 5ppm No3 and .06ppm Po4... and sucked out cyano weekly after it would coat my rocks again.

Well, everything looked good for awhile... but the dinos are increasing again on my sand bed! Well, the dinos are taking hold in areas of my sand where I had dumped sand back into my tank after "cleaning" it. During my siphoning of cyano out, I would usually get about 1-2 cups of sand as well... so I rinsed that sand with fresh water to lyse the dinos (since I would also siphon out areas with them in it as well)... and also treated that sand with some H2O2 to further kill dinos and cyano... then let it completely dry and then returned to the tank. Well, those areas are where the dinos are coming back! I think by cleaning the sand, it created a very sterile environment where the dinos had no competition; therefore, they are starting to flourish there. I wonder if after cleaning this sand, treating it by soaking it in a nitrifying bacteria solution (Microbacter7 + saltwater, etc...) and then returning it to the tank would keep the dinos from getting a foothold in the cleaned sand? Hmmm...

But, I think I'm going to give the Elegant Coral procedure a try. After so many months of battling with other treatments... I'm tired. A final decisive blow would be refreshing...
I have been mulling this over too for the spots I have amphidinium I have hanging on. I have not moved forward yet. I just got my skimmer back online. I did not want to attempt it without having the skimmer available.
I think that idea about soaking the the sand in bottled bacteria for awhile prior to placing it in back in the tank has merrit. Try it and share the results please. I have not been adding any sand back to my tank so could not tell it made a difference. And I wonder if placing that sand in an aquarium that was dino free to give it a chance to take on some life. But I think purchasing live sand from TBS would be a good idea too. But if I was wrong and all my expensive new LS became covered in dinos I would just roll over and die.
 
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taricha

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Well, the dinos are taking hold in areas of my sand where I had dumped sand back into my tank after "cleaning" it. During my siphoning of cyano out, I would usually get about 1-2 cups of sand as well... so I rinsed that sand with fresh water to lyse the dinos (since I would also siphon out areas with them in it as well)... and also treated that sand with some H2O2 to further kill dinos and cyano... then let it completely dry and then returned to the tank. Well, those areas are where the dinos are coming back! I think by cleaning the sand, it created a very sterile environment where the dinos had no competition;

That's a good interpretation, and maybe the correct one, but let me give you an alternative interpretation of the data. (I'm not sure which one I like better.)
By killing the dino cells and other organisms on the sand through freshwater and peroxide, then returning the sand you may have created rich organic matter in the form of the dead organisms still attached in parts to the sand. When @Dan_P and I try to grow these nuisances, we virtually always find organic matter as a key part of getting cyano (and maybe dinos) to grow.

Edit: no reason to either/or those two effects. You both killed the competition, and in so doing, provided dead organic matter to feed the first thing to recolonize the sand. Lucky dinos!
 
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Cruz_Arias

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That's a good interpretation, and maybe the correct one, but let me give you an alternative interpretation of the data. (I'm not sure which one I like better.)
By killing the dino cells and other organisms on the sand through freshwater and peroxide, then returning the sand you may have created rich organic matter in the form of the dead organisms still attached in parts to the sand. When @Dan_P and I try to grow these nuisances, we virtually always find organic matter as a key part of getting cyano (and maybe dinos) to grow.
We have also seen impacted and undigested organics/detritus as a source of nutrients for the dinos as well.

The solution, we came up with, was to re-activate sludge digesting/consuming bacteria and trigger a "feeding frenzy" response to help in the removal of trapped organics, similar to that in RAS in wastewater treatment.

Soaking in bacteria is good, provided that the substrate is aerobic and can sustain the bacterial seeding. Otherwise, the dead/weakened bacteria becomes another nutrient source as @taricha mentioned.
 

Victoria M

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That's a good interpretation, and maybe the correct one, but let me give you an alternative interpretation of the data. (I'm not sure which one I like better.)
By killing the dino cells and other organisms on the sand through freshwater and peroxide, then returning the sand you may have created rich organic matter in the form of the dead organisms still attached in parts to the sand. When @Dan_P and I try to grow these nuisances, we virtually always find organic matter as a key part of getting cyano (and maybe dinos) to grow.
So do not return the sand! No way to really clear the sand of these minute particles? Bleach washing? How much stirring and rinsing would it take to really clean the sand? Maybeway more than I want to think about. I threw my sand away the other day. I hosed and rinsed a few gallons of sand before I gave up on the rest of it. I figured it was not worth the work for the rest. But maybe I should just trash it all. Nice mesoflake stuff too. I might just set up a spare aquarium to give some life to this sand prior to to go to the tank.
 

Victoria M

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We have also seen impacted and undigested organics/detritus as a source of nutrients for the dinos as well.

The solution, we came up with, was to re-activate sludge digesting/consuming bacteria and trigger a "feeding frenzy" response to help in the removal of trapped organics, similar to that in RAS in wastewater treatment.

Soaking in bacteria is good, provided that the substrate is aerobic and can sustain the bacterial seeding. Otherwise, the dead/weakened bacteria becomes another nutrient source as @taricha mentioned.
Like do this in a separate container if you have removed your sand ?
 

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So do not return the sand! No way to really clear the sand of these minute particles? Bleach washing? How much stirring and rinsing would it take to really clean the sand? Maybeway more than I want to think about. I threw my sand away the other day. I hosed and rinsed a few gallons of sand before I gave up on the rest of it. I figured it was not worth the work for the rest. But maybe I should just trash it all. Nice mesoflake stuff too. I might just set up a spare aquarium to give some life to this sand prior to to go to the tank.
The regimen teaches how to keep the sand and turn it over and how to expose the unreacted organics in the sandbed. We stir a section of the sandbed, while actively bubbling, a third or a fourth at a time.

Aeration and continued aeration, with lower co2 air, will help you keep the system aerobic. That will also impact the sandbed, positively, up to 2 inches deep, approximately.
 
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taricha

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... And I love the idea of pre-treating the sand with a bacterial product to choose what colonizes it first. I wish I could tell you which bacterial product is most likely to discourage dino colonization.
But if we knew that, the hobby would be much farther along than it is now. :)
(Dr. Tim's Eco-Balance sure has an interesting description in this regard)
 

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That's a good interpretation, and maybe the correct one, but let me give you an alternative interpretation of the data. (I'm not sure which one I like better.)
By killing the dino cells and other organisms on the sand through freshwater and peroxide, then returning the sand you may have created rich organic matter in the form of the dead organisms still attached in parts to the sand. When @Dan_P and I try to grow these nuisances, we virtually always find organic matter as a key part of getting cyano (and maybe dinos) to grow.

Edit: no reason to either/or those two effects. You both killed the competition, and in so doing, provided dead organic matter to feed the first thing to recolonize the sand. Lucky dinos!

Dead organics may be the case, but those organics would have had to of had a pretty strong grip on the sands particles. I vigorously rinsed the sand many times after the peroxide with heavy agitation until the water ran clear.

I also started regular water changes and sand vacuuming again in the attempt to control the cyano. Then the dinos started to show!
 

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Dead organics may be the case, but those organics would have had to of had a pretty strong grip on the sands particles. I vigorously rinsed the sand many times after the peroxide with heavy agitation until the water ran clear.

I also started regular water changes and sand vacuuming again in the attempt to control the cyano. Then the dinos started to show!
The aragonite substrate is extremely porous. It used to hold life... so probability is that it has a high affinity to hold on to organics.
 

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... And I love the idea of pre-treating the sand with a bacterial product to choose what colonizes it first. I wish I could tell you which bacterial product is most likely to discourage dino colonization.
But if we knew that, the hobby would be much farther along than it is now. :)
(Dr. Tim's Eco-Balance sure has an interesting description in this regard)
I have some ATM Colony ordered and on the way (oredered the stuff from the ElegantCorals dino recipe). I'l give this a try with a couple dino sand patches prior to doing the EC dino recipe.
 

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I have some ATM Colony ordered and on the way (oredered the stuff from the ElegantCorals dino recipe). I'l give this a try with a couple dino sand patches prior to doing the EC dino recipe.
Using fresh air, low in co2, is very important. Plumb it to the Aqualifter and plumb an independent air intake for your skimmer.

Don't forget the airline valves to the wooden air stone and your skimmer air intake.
 

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A case study or studies on a random system would be interesting in conjunction with the analyzing of the method, right @taricha ?

Thoughts? Concerns?
 

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That's a good interpretation, and maybe the correct one, but let me give you an alternative interpretation of the data. (I'm not sure which one I like better.)
By killing the dino cells and other organisms on the sand through freshwater and peroxide, then returning the sand you may have created rich organic matter in the form of the dead organisms still attached in parts to the sand. When @Dan_P and I try to grow these nuisances, we virtually always find organic matter as a key part of getting cyano (and maybe dinos) to grow.

Edit: no reason to either/or those two effects. You both killed the competition, and in so doing, provided dead organic matter to feed the first thing to recolonize the sand. Lucky dinos!

Some ideas developed over the summer collaborating with @taricha

The nuisance benthic organism exists on a particular surface for a reason not by coincidence.

High density populations need a high concentration of food, greater than supplied by the water.

Bacteria are often closely associated with most organisms. The bacteria break down organic matter which a nuisance organism can use (N and P, for example) and the nuisance organism, typically photosynthetic, supplies organic carbon exudates to the bacteria. This mutualism may be the basis of the persistence of nuisance organism growth.

Particulate organic carbon can become concentrated, dissolved organic carbon cannot. High local concentrations of organic matter can easily be achieved. These fertile islands with the aid heterotrophic bacteria can become the home of most nuisance organisms. The high concentration of organisms can only last as long as fertile island does.
 

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