Analyzing a Bacterial Method for Dinoflagellates (and cyano?)

[Brew12]

Varous types of bacteria do react differently to different forms of carbon sources. IMO, vodka tends to cause the bacteria in Waste Away to react aggressively. Just an observation.

Absolutely, otherwise carbon dosing wouldn't work to reduce nitrates.

My observation from people using Waste Away has shown it reduces nitrates. My use of Vibrant and AOB's has not. In fact, I've seen nitrates increase when using Vibrant.

I'm confident that this method would not work trying to use Vibrant in place of Waste Away. I was only referring to the role that the One and Only (or other nitrifying bacteria) would play. AOB's are considered slow reproducers in the bacteria world.
However, it also can't be ruled out that the AOB's in One and Only do reproduce rapidly as has been shown in lab tests under specific conditions. If this is the case then the brand of AOB's used my also have a significant impact in the success of this method. The bottled bacteria do not all contain the exact same strains and different strains grow differently at different temperatures, pH, and sludge composition. So, this method may work with Dr Tims One and Only and not work with API Quick Start for example.

 

[CS Reefer]

I noticed when phosphates are zero/showing zero with standard titration tests, that nitrates held steady at an elevated level, using Waste Away, as well as other nitrifying bacteria strains. Could someone elaborate as to the effectiveness of similar strains, given a lack of phosphate please. The reverse seems to also be true, in relation to unmeasurable nitrates, and elevated phosphates.

 

[Brew12]

I noticed when phosphates are zero/showing zero with standard titration tests, that nitrates held steady at an elevated level, using Waste Away, as well as other nitrifying bacteria strains. Could someone elaborate as to the effectiveness of similar strains, given a lack of phosphate please. The reverse seems to also be true, in relation to unmeasurable nitrates, and elevated phosphates.

Phosphorus and nitrogen are cellular building blocks. Some organisms need more of each than others, and they have different ways to acquire them, but any bacterial based product will struggle without at least some of each.

This issue is very pronounced in QT/HT system with PVC pipe fittings and cycled with pure ammonia. If you keep cycling to 2ppm ammonia for a few weeks it will stop processing the ammonia until you add something like fish food to provide the bacteria the building blocks it needs.

 

[AngryOwl]

What a read...

 

[MickeyCT]

Wow! I go away for the weekend and tons of posts to read in this thread.

Mickey - Have you tried micro or nano bubbling at night with air from the outdoors. (fresh air) If it gets set up right is really helps with corals and some with ph stability or oxygenation of the water. Amazing the slime that corals release when bubbling occurs. The less CO2 in the system the less available to Dinos.

The method sort of resets the bacteria in the system. I have ran this once and it really knocked back the dinos but in 2 weeks I saw a few patches intermixed with my Diatoms and small patches GFA and GHA. After 5 weeks, I gave it another shot. I am only day 2 after the method, but no coral or fish loss. Sand is white and my nutrients are really low. Starting to raise them back up as corals recover method. If I see any rust/brown this time, I am running it one more time.

I have not tried this yet. I'll search for the instructions and give it a try. Thanks for bringing it up.

Having read some of @brandon429 threads on cleaning substrate, I was surprised to read that you had problems after replacing the sand bef.

Curious to see the work thread on it can you post it just to see what stands out. we like to check details like pre rinsing the new sand, rocks being cleared of detritus along with the sand being swapped (so that upwelling events wont feed invaders if present) and a few other details.

additionally, there are some rinses in the sand rinse thread that require hand guiding after the fact, follow up work. if the initial swap is left untended there are invaders that remass given enough time agreed to that as well. the #1 thing we try to impart is a bed that can be direct accessed and will not cause a recycle or loss to inhabitants as its worked as needed to suppress X, and then many times the rinse actually cures the issue anyway by sheer force of export.

When I replaced my sand bed, I relied on advice and help from "friends" who own and operate an aquarium maintenance company in town. Unfortunately, I had not read the thread(s) from brandon429. My bad for taking the easy road and I'm paying for it. ;Blackeye

 

[taricha]

And with regard to carbon dosing, is the thinking that ethanol will help heterotrophic bacteria consume organic matter that it would not ordinarily consume in the absence of a simple carbon source?

...or does it just grow lots of bacteria?

Hey, have you looked at WasteAway in a test tube in any manner, say, turbidly vs mg of organic material? Turbidity vs type of organic matter?

Wanted to revisit this as I got somewhat of a result with my aged grunge from an algae tank.

The waste away alone created some noticeable cloudiness, and waste away + vodka significantly more cloudiness. Pristine by seachem (another bacterial "grunge eater" product) produced no cloudy water bloom, it remained as clear as the control tubes. Additional tubes of vibrant, vibrant+vodka, and vodka only also remained clear.

Also the majority of the tubes stayed the same shade of light fluffy brown, but the two Waste-Away treatments started to turn color to the gray/black appearance that I associate with matter becoming anoxic. The vodka only treatment also turned gray.
Aside: I don't know why already dead material would change colors to gray/black due to low oxygen. Seems like being dead already, it wouldn't get more dead.

[The waste-away turned grayish, and the waste away + vodka had a stronger color change toward gray/black.]

My conclusions:
Waste-Away activity alone is sufficient to create cloudy water (bacterial bloom) in the presence of old otherwise stable grunge.
Ethanol supercharges this bacterial bloom process, but is not sufficient to create it alone.
Not every "grunge-eater" bottled bacteria is capable of creating a cloudy water bloom event from this old mostly stable material.
The bloom is oxygen-hungry and creates a low oxygen environment with or without ethanol addition but more intense with it.

These observations are all supportive of the proposed role of Waste-Away and ethanol as described in the method (except that the cloudiness is probably not from the nitrifying bacteria. The Waste-away is sufficient to make water cloudy)

Final comment: I was surprised to find that I can clearly microscopically distinguish the bacteria in Waste-Away, One and Only, and Vibrant. They are quite different in appearance and behavior. Hopefully I'll be able to say something about how the cloudy water bloom bacteria do or don't resemble the waste-away that fuels the bloom.

edit: video - Waste Away under microscope

 

[Dan_P]

...or does it just grow lots of bacteria?

Wanted to revisit this as I got somewhat of a result with my aged grunge from an algae tank.

The waste away alone created some noticeable cloudiness, and waste away + vodka significantly more cloudiness. Pristine by seachem (another bacterial "grunge eater" product) produced no cloudy water bloom, it remained as clear as the control tubes. Additional tubes of vibrant, vibrant+vodka, and vodka only also remained clear.

Also the majority of the tubes stayed the same shade of light fluffy brown, but the two Waste-Away treatments started to turn color to the gray/black appearance that I associate with matter becoming anoxic. The vodka only treatment also turned gray.
Aside: I don't know why already dead material would change colors to gray/black due to low oxygen. Seems like being dead already, it wouldn't get more dead.

[The waste-away turned grayish, and the waste away + vodka had a stronger color change toward gray/black.]

My conclusions:
Waste-Away activity alone is sufficient to create cloudy water (bacterial bloom) in the presence of old otherwise stable grunge.
Ethanol supercharges this bacterial bloom process, but is not sufficient to create it alone.
Not every "grunge-eater" bottled bacteria is capable of creating a cloudy water bloom event from this old mostly stable material.
The bloom is oxygen-hungry and creates a low oxygen environment with or without ethanol addition but more intense with it.

These observations are all supportive of the proposed role of Waste-Away and ethanol as described in the method (except that the cloudiness is probably not from the nitrifying bacteria. The Waste-away is sufficient to make water cloudy)

Final comment: I was surprised to find that I can clearly microscopically distinguish the bacteria in Waste-Away, One and Only, and Vibrant. They are quite different in appearance and behavior. Hopefully I'll be able to say something about how the cloudy water bloom bacteria do or don't resemble the waste-away that fuels the bloom.

edit: video - Waste Away under microscope

You and your work are far more entertaining than the PBS series NOVA.

Some questions on the nature of the grunge.
If you filtered or centrifuged this stuff and then dried the solid, is there much mass left? Would it burn?
I wonder what the element make up of the grunge is?
Would cyanobacteria grow on it?
Does grunge consume hydrogen peroxide?
Is grunge soluble at high or low pH?

Bacteria questions
Do these grunge eating bacteria consume the slime produced by dinoflagellates?
Do these bacteria consume biofilms? Would these bacteria clean old substrate (still trying to imagine how to observe or measure this)
What happens to nitrate and phosphate during grunge digestion? Does phosphate become limiting? NO3?
Would these bacteria consume ground up GHA? Wondering if they need an ideal C:N ratio to live?

I can buy these bacteria and try to answer these questions (though cyanobacteria studies still consuming much of my lab budget) but I don’t have a source of grunge. Remind me, is grunge material sucked out of the substate or the fluff in the sump?

 

[Dan_P]

...or does it just grow lots of bacteria?

Wanted to revisit this as I got somewhat of a result with my aged grunge from an algae tank.

The waste away alone created some noticeable cloudiness, and waste away + vodka significantly more cloudiness. Pristine by seachem (another bacterial "grunge eater" product) produced no cloudy water bloom, it remained as clear as the control tubes. Additional tubes of vibrant, vibrant+vodka, and vodka only also remained clear.

Also the majority of the tubes stayed the same shade of light fluffy brown, but the two Waste-Away treatments started to turn color to the gray/black appearance that I associate with matter becoming anoxic. The vodka only treatment also turned gray.
Aside: I don't know why already dead material would change colors to gray/black due to low oxygen. Seems like being dead already, it wouldn't get more dead.

[The waste-away turned grayish, and the waste away + vodka had a stronger color change toward gray/black.]

My conclusions:
Waste-Away activity alone is sufficient to create cloudy water (bacterial bloom) in the presence of old otherwise stable grunge.
Ethanol supercharges this bacterial bloom process, but is not sufficient to create it alone.
Not every "grunge-eater" bottled bacteria is capable of creating a cloudy water bloom event from this old mostly stable material.
The bloom is oxygen-hungry and creates a low oxygen environment with or without ethanol addition but more intense with it.

These observations are all supportive of the proposed role of Waste-Away and ethanol as described in the method (except that the cloudiness is probably not from the nitrifying bacteria. The Waste-away is sufficient to make water cloudy)

Final comment: I was surprised to find that I can clearly microscopically distinguish the bacteria in Waste-Away, One and Only, and Vibrant. They are quite different in appearance and behavior. Hopefully I'll be able to say something about how the cloudy water bloom bacteria do or don't resemble the waste-away that fuels the bloom.

edit: video - Waste Away under microscope

Just watched the video. These bugs would be countable. An important consideration in growth studies.

 

[taricha]

Final comment: I was surprised to find that I can clearly microscopically distinguish the bacteria in Waste-Away, One and Only, and Vibrant. They are quite different in appearance and behavior. Hopefully I'll be able to say something about how the cloudy water bloom bacteria do or don't resemble the waste-away that fuels the bloom.

video - Waste Away under microscope

Ok, after looking carefully at the cloudy water itself....
It does not much resemble Waste-Away (or any other bacterial product). It's a complete zoo of multiple shapes and sizes of bacteria moving in totally different ways and speeds.
Video (2min) of cloudy water bacteria:


Check it out at 0:35, 1:20 and 1:45 for a flavor.
(turn the resolution to max to catch all the tiny zooming streaks)
Comparing to the waste-away video from my previous post, and other info from previous post, here's my tentative interpretation. (feel free to shoot it down):
Waste-Away is the primary cause of the cloudy water bloom. It supports it and likely participates in it, but the cloudy water is by no means primarily waste-away. It's a wide variety of different bacteria benefitting from the conditions waste-away/ethanol created.
If that's true, then pairing the waste-away with another desirable bacterial blend makes a lot of sense, as it may give use some amount of control over what we are growing a ton of in the cloudy water.

 

[taricha]

You and your work are far more entertaining than the PBS series NOVA.

Some questions on the nature of the grunge.
If you filtered or centrifuged this stuff and then dried the solid, is there much mass left? Would it burn?
I wonder what the element make up of the grunge is?
Would cyanobacteria grow on it?
Does grunge consume hydrogen peroxide?
Is grunge soluble at high or low pH?

Do these grunge eating bacteria consume the slime produced by dinoflagellates?
Do these bacteria consume biofilms? Would these bacteria clean old substrate (still trying to imagine how to observe or measure this)
What happens to nitrate and phosphate during grunge digestion? Does phosphate become limiting? NO3?
Would these bacteria consume ground up GHA? Wondering if they need an ideal C-N-P ratio to live?

I can buy these bacteria and try to answer these questions (though cyanobacteria studies still consuming much of my lab budget) but I don’t have a source of grunge. Remind me, is grunge material sucked out of the substate or the fluff in the sump?

Grunge:
was siphoned off the top of the bed of a neglected macroalgae tank. My bet is that this contains a lot of algal cell walls, I'd think quite a bit of cellulose in the fluff. I suspect that because the tank has been mostly just growing macroalgae. And I've also observed dead cheato strands in test tubes of tank water, and although they grew cyano, diatoms, etc all around them, the skeleton chaeto strands themselves were not broken down in months. So maybe this fluffy stuff that hasn't changed much in months shares some similar content. I'll see if it's burnable.

I did a whole microscope and pigment analysis of the grunge before I started. It is microscopically unrecognizable debris that supports a sparse collection of strands of derbesia, cyano filaments, and diatoms - some ciliates and tiny pods in it as well. The extracted pigments tell the same story as the microscope.

Bacteria:
I think it would love dino mucus. Trying to grow enough dinos to check this.
This old grunge comes from a tank where all available N and P is already thoroughly consumed by macroalgae. So for the waste-away to trigger a bloom in these conditions it has to be liberating nutrients that were previously locked up.

 

[Dan_P]

Grunge:
was siphoned off the top of the bed of a neglected macroalgae tank. My bet is that this contains a lot of algal cell walls, I'd think quite a bit of cellulose in the fluff. I suspect that because the tank has been mostly just growing macroalgae. And I've also observed dead cheato strands in test tubes of tank water, and although they grew cyano, diatoms, etc all around them, the skeleton chaeto strands themselves were not broken down in months. So maybe this fluffy stuff that hasn't changed much in months shares some similar content. I'll see if it's burnable.

I did a whole microscope and pigment analysis of the grunge before I started. It is microscopically unrecognizable debris that supports a sparse collection of strands of derbesia, cyano filaments, and diatoms - some ciliates and tiny pods in it as well. The extracted pigments tell the same story as the microscope.

Bacteria:
I think it would love dino mucus. Trying to grow enough dinos to check this.
This old grunge comes from a tank where all available N and P is already thoroughly consumed by macroalgae. So for the waste-away to trigger a bloom in these conditions it has to be liberating nutrients that were previously locked up.

Thanks for the details.

I have a hypothesis that anything that hangs around an aquarium for awhile is near or at the end of its nutritional value. Grunge might serve as a habitat but not food, unless it is serving as a trap for particulate organic carbon. I never got around to test the idea, hence my earlier questions.

A second thought comes to mind about where the added bacteria is doing it’s work. I have a notion that most bacterial digestion happens on surfaces. The water is poor place to eat. If a bacteria blooms occurs in the water, it occurs because something was added to the water, in this case ethanol. While that might be useful for nitrate reduction, I now wonder whether that will turn out to be a sideshow in the grunge reduction saga because the real work must be done on or under the surface. I guess it is possible that the water is being loaded with bacterial waste from the digestion of the grunge. This might be testable.

You can see why your posts surpass NOVA in being thought provoking.

 

[CS Reefer]

It would be interesting to see a similar experiment, using freshly mixed salt water, with the addition of measured nitrates and phosphates. Does the bloom happen without adding the debris (since the constituents are somewhat varied and unknown)?

 

[Dan_P]

It would be interesting to see a similar experiment, using freshly mixed salt water, with the addition of measured nitrates and phosphates. Does the bloom happen without adding the debris (since the constituents are somewhat varied and unknown)?

This might help sort things out a bit.

 

[taricha]

See, that's the sort of experiment I wouldn't think to do because I already know the answer.

(Then when I do it anyway, it doesn't turn out like I expected )

I'm thinking yes. If you add N and P and a carbon dose you'll get a bloom regardless of bottled bacteria added or not.

At the moment I'm thinking about how to tell if the added ethanol actually enables the Waste Away to consume more grunge or if it just intensifies the bacterial growth allowing you to export more efficiently (skimmer/siphon).
Working on quantifying grunge consumed by bacteria.

 

[Dan_P]

See, that's the sort of experiment I wouldn't think to do because I already know the answer.

(Then when I do it anyway, it doesn't turn out like I expected )

I'm thinking yes. If you add N and P and a carbon dose you'll get a bloom regardless of bottled bacteria added or not.

At the moment I'm thinking about how to tell if the added ethanol actually enables the Waste Away to consume more grunge or if it just intensifies the bacterial growth allowing you to export more efficiently (skimmer/siphon).
Working on quantifying grunge consumed by bacteria.

A simple but slow method to study grunge might be with aerobic digestion. The setup is simple too. A capped bottle fitted with a bubbler. Aerobic digestion will produce CO2, NH3 and PO4, all easily measured. In time the bacteria will include NH3 oxidizers and nitrate will be produced. Digestion time could take weeks. Aquarium water would serve as the medium to which bacteria and grunge are added. Aquarium water would serve as a control. You may be surprised at the amount of nitrate produced with nothing added, not even WasteAway.

I feel that you might get useful information about the impact of ethanol addition to grunge digestion outcome. CO2 production, NH3/NO3 and PO4 accumulation trends are the data that I am thinking might be useful. Alkalinity and pH might be useful too.

Biological oxygen demand would be an even simpler technique to characterize grunge and a measurement would take only a week.

 

[taricha]

A simple but slow method to study grunge might be with aerobic digestion.

I set up a similar thing. Slow bubbling grunge in glass tube.
Turn off bubbling to measure grunge level.

Attempt to use Waste Away (then add ethanol) to measurably reduce the amount of grunge.

Your suggestion would probably tell us much more though.

 

[Dan_P]

I set up a similar thing. Slow bubbling grunge in glass tube.
Turn off bubbling to measure grunge level.

Attempt to use Waste Away (then add ethanol) to measurably reduce the amount of grunge.

Your suggestion would probably tell us much more though.

Oh you clever boy! Great idea.

I hope you are thinking of peeking at the water chemistry now and then, O2 too, and measuring the CO2 concentration in a volume of the off gas. I’m stuck obsessively studying cyanobacteria, otherwise, I’d collect some numbers along with you

 

[CS Reefer]

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”

 

[Dan_P]

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”

Me too!

 

[taricha]

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.