Analyzing a Bacterial Method for Dinoflagellates (and cyano?)

[taricha]

I've seen this method posted about and tried here and there. It has more than enough success to warrant its own thread and a closer look. Failures have been of the form - it bloomed too hard and killed things. Because it relies almost entirely on the invisible activity of bottled bacterial products, it's hard to evaluate the claims about what it does and how it works.
That's what I'd like to take a closer look at. Hope to get some knowledgable eyeballs in this thread who can help understand what's going on, see if the mechanics are described properly and maybe evaluate some of the points.

This is from Elegant Corals

And here is a youtube video where the author, Cruz Arias discusses the method in more detail (from 5 min to 29 min).

The overall theme is two bacterial blends: one a grunge-eater remineralizes debris particles into inorganic nutrients, and a second bacterial actor consumes the nutrients, multiplies forming a bacterial bloom and is removed by skimming.

Here as I understand it - are the details of the mechanics of the process:

1. Aerate the system using a low-CO2 air input with very fine bubbles to drive off as much CO2 as possible:
   a. limit encouragement to dino growth.
   b. [author claim] some dinos die from CO2 deprivation caused by aggressive low-CO2 aeration
   c. prevent oxygen deprivation from the large bacterial bloom to come.
   d. Bacterial activity later will generate CO2, and aggressive bubbling drives this away
2. Dr Tim's Waste Away - consume debris and organic matter and remineralize it to N and P
   a. may also break down dino mucus mats, disrupt dino bacterial partners
   b. [author claim] there is no comparable substitute for Waste-Away(!)
3. ATM Colony (or Dr Tim One and Only or other tank starter nitrifying bacteria)
   a. consume the liberated nutrients from the Waste-Away breakdown process
   b. may multiply as nutrients are consumed and serve as nutrient export through skimming (do nitrifying bacteria multiply quickly?)
   c. [author claim] the nitrifier bacteria blend are the ones that get exported through skimming
   d. may also serve to ensure ammonia generated anywhere in tank is immediately processed
4. Vodka as a large carbon source from beginning
   a. boost and accelerate the bacterial processes
   b. help the Waste-Away digest particulates?
   c. help grow the nitrifiers?
   d. [author claim] most tanks tend to be carbon limited unless specifically dosed
5. Turn off Skimming - early on, trying to create a bacterial bloom
6. Stir 1/3 of sand bed
   -bring bacterial population in contact with pockets of debris and dino matter packed in the sandbed
   -agitation of dinos (cyano) creates a clumping response, making them easier to remove
7. Manually siphon any visible clumps of dinos/cyano - direct cell removal is always a good step forward
8. Turn on skimming - at day 3, if the bloom is pushing pH too low (<7.6)
   a. day 4, skimmer gets turned on regardless, and the bacterial export begins
9. Hydrogen peroxide (common 3%) - at day 3 if bloom is too aggressive, add h2o2.
   a. day 4, h2o2 gets dosed to begin cutting down bacterial activity
   b. peroxide slow the bacterial activity or kill them?

 

[Cory]

Ive always said cyano and dinoflagellets come from rotting organics. Possibly all thats needed is dr tims waste away in this method.

 

[Dan_P]

The overall description sounds like an aerobic digester used in sewage remediation. Because bacteria use organic carbon both for energy and biomass creation, there is going to be a stoichiometric excess of nitrogen and phosphorous and remineralization of N and P occurs. Addition of organic carbon to sewage is not uncommon. The aeration makes sense if you want to minimize the chance of the digester becoming anaerobic.

When running an aerobic digester though, you must have a sense of the organic load you want consumed snd the CO2 production and oxygen consumption rates. This methodology provides no guidance or analytical method to help you monitor the progress the digestion in your system. As a guiding principle, it is not a good idea to add things to your system and not understand what they are doing.

And then there is the all important discussion of success rate. How often does this procedure work? Where are the statistics? In god we trust, everyone else show me the data.

 

[AngryOwl]

Thanks for making this thread! I'm currently trying this method. On day 2 the water was noticeably cloudy and day 3 I lost my pintail wrasse. I was pulling air from inside thinking it would be enough. I've since moved 2 airpumps outside so a lack of oxygen at this point seems unlikely. I spoke with elegant corals about it and it was stated there was not enough oxygen and the bacteria basically died. He said it was a perfect environment for my new dino pets though.

This method is still inconclusive to me and lacks a key faq, as well as a deeper understanding of what the above poster mentions.

Anyways I've restarted it and I'll report back what I see.

 

[AngryOwl]

After reading Dans comments above previously, it really got me thinking about how reckless this method actually is. A lack of official research and an FAQ is even more concerning.

If you have fish, this method is reckless IMO. The method calls for a 'controlled' bacteria bloom. One that will consume an unknown amount of oxygen. What it does not account for is the life already in your tank (fish) that require oxygen. Do the same instructions apply if you have 25 fish, compared to having 5 in the same tank size? The obvious answer is no. Thus you risk suffocating your fish.

I suppose this all comes down to the air pump you choose to pull in fresh air. I would see this as less reckless if you use the largest pump you can get.

We also have to examine this method comes from the sewage industry method... where there is no regard for life other than bacteria. IMO fish should be removed and then go ahead and do the treatment.

 

[taricha]

@AngryOwl thanks for your comments. I'd like to know what exactly were the aeration procedures, given that they weren't enough?

 

[taricha]

Let me narrow things down this way.

Of my little numbered list in the first post, these are the ones I am pretty sure are correct, or do what they are expected.
1,a,d.
2.
3,a,d.
4,a,d.
5.
6,a,b.
7.
8.

So what does that leave?
1b - some dinos die from CO2 being cut off through low-CO2 aeration.
I can't see how this would be true. Slower dino growth if equillibrium with atmospheric CO2, but not death of cells within a day or two. Not possible is it?

1c - aeration to prevent oxygen deprivation from the large bacterial bloom to come.
Obviously we need to talk about this. The method seems to have a lot of control mechanisms, yet some people still get livestock death from low O2 during the bloom. What level of aeration would actually protect inhabitants?

2a Waste Away - may also break down dino mucus mats,
I need to dig up a paper on relation between dino toxins and bacteria, but I recall suggestion that bacteria populations could increase or remove the toxins of a dino bloom. The mucus mats may be entirely digestible by the right bacteria. Waste-away seems like a good candidate.

2b - there's no comparable substitute for Waste-Away(!)
Really?!? that's extraordinary if true. I have no idea if it's true.

3b - Tank Starter Nitrifying (denitrifying?) bacteria multiply rapidly as nutrients liberated by Waste-Away
Why does a tank take so long to cycle if nitrifiers are capable of explosive growth under these conditions?

3c - the nitrifier bacteria blend are the ones that get exported through skimming
Probably true. I don't think the waste away is forming a large population anywhere other than the substrate. So cloudiness in the water may be the nitrifying bacteria?

4b Vodka - help the Waste-Away digest particulates?
As @Dan_P says there's good reason to expect this is true and that Carbon limitation is an impediment to digestion of the particulates. This is a really helpful thing to know. No question on this point. I just put it here to talk about it for emphasis.

4c Vodka help grow the nitrifiers?
This one I dunno. Does having organic carbon around make nitrification (denitrification) easier? more productive?

9a day 4, h2o2 gets dosed to begin cutting down bacterial activity
9b peroxide slow the bacterial activity or kill them?
This method uses peroxide like a brake pedal (and vodka like a gas pedal) to keep the bacterial bloom within some guardrails. I've never heard of peroxide used like this, to check a bacterial population. Can this really work?


@Dan_P also brought up the idea that this method may be sensitive to the amount of grunge there is available to feed the bloom, and it may not be well controllable or proceed to completion without that knowledge.

Ive always said cyano and dinoflagellets come from rotting organics.

I agree on cyano. It seems deeply tied to the debris in our tanks. Dinos may be more versatile in how they get their nutrients. But it's certainly a big factor.

 

[taricha]

I did a little experiment to get some data on O2 and CO2 during bacteria bloom. Take this data with a grain of salt. None of these measuring devices are meant for aquarium use, and my calibrations could be dodgy etc. Please call me out if any of this data seems implausible.

In the afternoon, I started a bacterial bloom with Dr. Tim's Waste Away and added carbon Vodka & vinegar. (I did not start aeration at this point.)

The next mornign the water was cloudy so I grabbed a dissolved oxygen probe and measured 60% (of max dissolved oxygen). Since that was at the top of the tank, I sampled from bottom of tank, a few inches off the sand and got 49%.
Wanting to see if it could be pushed further, I added a little more Waste-Away and carbon source, and stirred the sandbed - blew around a lot of gunk. (I never vacuum my sand.)
The dissolved oxygen stayed around 60% and didn't really move over the next half-hour, so I turned off the main tank lights to see if things would get sketchy. Yep - in a hurry. Less than an hour, top of my tank had dropped from 60% to 19% oxygen.
Lights on, aeration on got it back up to 30% where it basically stayed level. I turned my skimmer on (pulling from outside) with the collection cup off for as much aeration as possible. couple of hours later, the oxygen stabilized but never rebounded above 32%. I made sure to turn my sump lights to always on. If my entire system went dark for any length of time, then I'm not sure even a ton of aeration would keep things from going badly. All this was under 24hrs.

next CO2. The bacteria consuming gunk ought to be producing some CO2. It's not a dissolved CO2 probe, it measures CO2 in the air. In the room it's 500-600ppm. To compare water CO2 levels, I took 100mL water put in a 250mL bottle and shake it vigorously for ~1min to equalize gasses in the bottle. Then measure the air in the bottle for CO2. distilled water and saltwater from a tank that's not being messed with both gave readings close to room levels ~600-700ppm CO2.
Water from the tank during the bloom gave me anywhere from 1800ppm CO2 when not aerated, down to 1200ppm when aerated as strongly as I can manage.
so 3x normal CO2 levels and aerating the heck out of it, I could get it down to 2x normal levels. The bacteria are producing CO2 rapidly, and the best aeration I can achieve could not bring it down to near normal ambient levels.

I also measured pH after all this and got 7.55 - this is below the level (7.6pH) that the author of the guide says you should use peroxide to start putting bloom in check.

Some aquarium oxygen background reading for those like me who hadn't tracked aquarium dissolved oxygen.
Eric Borneman: The Need to Breathe Part 1
Part 2
Part 3
(part 3 most applicable. Data for clownfish tank with no skimmer at night, and creating bacterial bloom in a stagnant tank using live rock show results sort of like what I saw).

 

[MrAlpha]

What was the PH prior to the bloom? In other words, how far down did the Bloom drive the PH?

 

[Codiak]

Is this like homemade Micro Bacter7 and Reef BioFuel? Which is so much safer and simpler.

 

[taricha]

What was the PH prior to the bloom? In other words, how far down did the Bloom drove the PH?

Let me try to get some data that'll be better than me just guessing.
(Might be imagining, but I think at a given Alk - only CO2 moves the pH)

 

[AngryOwl]

I did a little experiment to get some data on O2 and CO2 during bacteria bloom. Take this data with a grain of salt. None of these measuring devices are meant for aquarium use, and my calibrations could be dodgy etc. Please call me out if any of this data seems implausible.

In the afternoon, I started a bacterial bloom with Dr. Tim's Waste Away and added carbon Vodka & vinegar. (I did not start aeration at this point.)

The next mornign the water was cloudy so I grabbed a dissolved oxygen probe and measured 60% (of max dissolved oxygen). Since that was at the top of the tank, I sampled from bottom of tank, a few inches off the sand and got 49%.
Wanting to see if it could be pushed further, I added a little more Waste-Away and carbon source, and stirred the sandbed - blew around a lot of gunk. (I never vacuum my sand.)
The dissolved oxygen stayed around 60% and didn't really move over the next half-hour, so I turned off the main tank lights to see if things would get sketchy. Yep - in a hurry. Less than an hour, top of my tank had dropped from 60% to 19% oxygen.
Lights on, aeration on got it back up to 30% where it basically stayed level. I turned my skimmer on (pulling from outside) with the collection cup off for as much aeration as possible. couple of hours later, the oxygen stabilized but never rebounded above 32%. I made sure to turn my sump lights to always on. If my entire system went dark for any length of time, then I'm not sure even a ton of aeration would keep things from going badly. All this was under 24hrs.

next CO2. The bacteria consuming gunk ought to be producing some CO2. It's not a dissolved CO2 probe, it measures CO2 in the air. In the room it's 500-600ppm. To compare water CO2 levels, I took 100mL water put in a 250mL bottle and shake it vigorously for ~1min to equalize gasses in the bottle. Then measure the air in the bottle for CO2. distilled water and saltwater from a tank that's not being messed with both gave readings close to room levels ~600-700ppm CO2.
Water from the tank during the bloom gave me anywhere from 1800ppm CO2 when not aerated, down to 1200ppm when aerated as strongly as I can manage.
so 3x normal CO2 levels and aerating the heck out of it, I could get it down to 2x normal levels. The bacteria are producing CO2 rapidly, and the best aeration I can achieve could not bring it down to near normal ambient levels.

I also measured pH after all this and got 7.55 - this is below the level (7.6pH) that the author of the guide says you should use peroxide to start putting bloom in check.

Some aquarium oxygen background reading for those like me who hadn't tracked aquarium dissolved oxygen.
Eric Borneman: The Need to Breathe Part 1
Part 2
Part 3
(part 3 most applicable. Data for clownfish tank with no skimmer at night, and creating bacterial bloom in a stagnant tank using live rock show results sort of like what I saw).

This is awesome analysis - 100% scientific or not, it provides insight. What I find interesting is your inside air ambient CO2 levels. The author claims homes tend to average around 1000ppm CO2, while outside averages 450ppm CO2. This is the reasoning behind pulling outside air. However, if you're around 500-600, it seems negligible to me.

When I began the method I had my air pump (tetra whisper 20) pulling air from under the stand. My stand is fully wide open FYI. I live in a 2700sqft home, 2 owls ('humans') that work during the day, 3 dogs, and a cat... the home also has tall ceilings and the A/C unit pulls fresh air into the home because foam insulation is used. I mention all this to say I 'feel' it is unlikely my home would be in the 1000ppm CO2 range.

What I can only speculate is the air pump was not enough to keep up with the bacteria demand. -- However! -- When I added a 2nd air pump and placed both outside I noticed similar results in my PH. For instance, the PH would dip slightly below 7.5 at night with 1 pump inside AND with 2 pumps outside. Coincidence? I think not.

While I've had no additional fish loss from this 2nd round - I haven't seen any difference in the tanks PH because I'm pulling outside air. I will say however my tank is looking better. Today is day 7 and the sand bed is looking dino free and my rocks are looking cleaner. I cannot say with 100% certainty this method produced the results solely though.

What would be another interesting experiment is seeing how much air is required to keep O2 levels during the bloom 'safe'. On a sample tank this would obviously need to be scaled up. For instance if it takes 1 whisper 20 pump to keep O2 safe within a 50g tank - it would be safe to say, maybe, that you would need 4 times the power for a 200g tank.

I do not appreciate the 'one size fits all' approach the author has taken without providing caveats to the key nuances within different tanks or any supporting scientific research.

 

[brandon429]

we should instantly turn this into a live works thread agreed on looking into method. w try to locate and aim some participants this way. large tankers relish a water control system / no headaches probiotics lets go

we need to start w pics that show progression to find patterns as the thread adds pages

 

[taricha]

we should instantly turn this into a live works thread agreed on looking into method. w try to locate and aim some participants this way. large tankers relish a water control system / no headaches probiotics lets go

we need to start w pics that show progression to find patterns as the thread adds pages

Let's pump the brakes on that. As I get into this, the more I think there are misunderstandings of what the bacteria are doing. (Or at least imprecise descriptions)
The author's description of what the bacteria do and Dr Tim's description of his own products seem in conflict.
More details later.

 

[brandon429]

how are we going to know if the claims are valid wo work examples for patterns. figured it would be neat if ten participants showed up only using the written method above, to track outcomes

 

[taricha]

This is awesome analysis - 100% scientific or not, it provides insight. What I find interesting is your inside air ambient CO2 levels. The author claims homes tend to average around 1000ppm CO2, while outside averages 450ppm CO2. This is the reasoning behind pulling outside air. However, if you're around 500-600, it seems negligible to me.

Important caveat!
This tank is in a school lab that's been vacant virtually all summer.
During year, room CO2 on similar meters shows values in the thousands.
In a lived-in space, inside/outside CO2 is not negligible difference.

 

[taricha]

how are we going to know if the claims are valid wo work examples for patterns. figured it would be neat if ten participants showed up only using the written method above, to track outcomes

Then I'd say, proceed only if you can actively monitor pH or O2 (and maybe Ammonia. I haven't looked at that yet.)
The instructions alone don't guard against unhealthy tank conditions.

 

[brandon429]

oh I see, concern about losses I see. not trying to nuke tanks /check. good write up and idea, I saw it a few mos ago and wondered about it

 

[Dan_P]

I did a little experiment to get some data on O2 and CO2 during bacteria bloom. Take this data with a grain of salt. None of these measuring devices are meant for aquarium use, and my calibrations could be dodgy etc. Please call me out if any of this data seems implausible.

In the afternoon, I started a bacterial bloom with Dr. Tim's Waste Away and added carbon Vodka & vinegar. (I did not start aeration at this point.)

The next mornign the water was cloudy so I grabbed a dissolved oxygen probe and measured 60% (of max dissolved oxygen). Since that was at the top of the tank, I sampled from bottom of tank, a few inches off the sand and got 49%.
Wanting to see if it could be pushed further, I added a little more Waste-Away and carbon source, and stirred the sandbed - blew around a lot of gunk. (I never vacuum my sand.)
The dissolved oxygen stayed around 60% and didn't really move over the next half-hour, so I turned off the main tank lights to see if things would get sketchy. Yep - in a hurry. Less than an hour, top of my tank had dropped from 60% to 19% oxygen.
Lights on, aeration on got it back up to 30% where it basically stayed level. I turned my skimmer on (pulling from outside) with the collection cup off for as much aeration as possible. couple of hours later, the oxygen stabilized but never rebounded above 32%. I made sure to turn my sump lights to always on. If my entire system went dark for any length of time, then I'm not sure even a ton of aeration would keep things from going badly. All this was under 24hrs.

next CO2. The bacteria consuming gunk ought to be producing some CO2. It's not a dissolved CO2 probe, it measures CO2 in the air. In the room it's 500-600ppm. To compare water CO2 levels, I took 100mL water put in a 250mL bottle and shake it vigorously for ~1min to equalize gasses in the bottle. Then measure the air in the bottle for CO2. distilled water and saltwater from a tank that's not being messed with both gave readings close to room levels ~600-700ppm CO2.
Water from the tank during the bloom gave me anywhere from 1800ppm CO2 when not aerated, down to 1200ppm when aerated as strongly as I can manage.
so 3x normal CO2 levels and aerating the heck out of it, I could get it down to 2x normal levels. The bacteria are producing CO2 rapidly, and the best aeration I can achieve could not bring it down to near normal ambient levels.

I also measured pH after all this and got 7.55 - this is below the level (7.6pH) that the author of the guide says you should use peroxide to start putting bloom in check.

Some aquarium oxygen background reading for those like me who hadn't tracked aquarium dissolved oxygen.
Eric Borneman: The Need to Breathe Part 1
Part 2
Part 3
(part 3 most applicable. Data for clownfish tank with no skimmer at night, and creating bacterial bloom in a stagnant tank using live rock show results sort of like what I saw).

Nice investigation! Which system did you use?

What oxygen meter are you using?

What are you using to measure CO2?

I wonder about the oxygen level dropping to 19% but never recovering to above 30%.

Headspace analysis is what you did. Further reading might help design a continuous monitoring experimental set up.

Borneman neglected to measure the air flow when aerating the system with an air stone and skimmer. That is important data for comparing the two methods as would bubble size. Also, I did not see a discussion on scaling up gas exchange in aquaria (I read the articles quickly). All this would be important in determining how to keep your system oxygenated and minimizing CO2 build up.

You should do some figurin’ to see if your results are within the bounds of possibility. Here are some questions that might be useful?

What is the concentration of CO2 in the system when the pH drops from X to 7.55?

Would you expect the headspace above distilled water and take water to be approximately the same? Show work

What concentration of CO2 in seawater would be in equilibrium with 1800 ppm CO2 in the headspace?

If all the carbon in the vodka and vinegar is converted to CO2 and did not leave the tank, what would be the concentration in the system?

How much oxygen is consumed in converting all the added ethanol and vinegar to CO2?

 

[taricha]

Nice investigation! Which system did you use?
What oxygen meter are you using?
What are you using to measure CO2?

My main 70 gal system.
Vernier Dissolved Oxygen Probe and CO2 gas sensor

I think I recorded enough info, and can probably track down enough reading material to be able to work out most of that stuff

What was the PH prior to the bloom? In other words, how far down did the Bloom drive the PH?

Two meters both said a drop of about ~0.45 to 0.50pH when O2 was down around 20% of max.
The better meter (newer Hanna) said 8.12->7.65