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BBA some exploratory experiments

7K views 31 replies 9 participants last post by  mistergreen 
#1 · (Edited)
I am recently experiencing a renewed bloom of BBA in my aquarium. So now that I have the raw material I can plan the experiments.

General details
After I pick mature BBA from the aquarium I put them into small covered glass containers. They will be monitored and photographed. I will consider them growing if they are increasing in size or new algae colonies are observable. I will consider them "dead" if the algae is white-transparent or red and does not recover after 1 week. The light will be coming from a west window ( spectrum and amount does not matter for these experiments). All the water has a starting concentration of :
NO3 15 ppm, PO4 2.7 ppm, 11.2 ppm, 0.4 ppm Mg. Tap water has a reported GH 13, KH 11.


Controls: 2 bottles with tap water exposed to light
Experiment 1: Simply keep the algae samples in the dark for 3,5,10,15,20 days. After the darkness period re-expose to normal light and observe.

Experiment 2: Put algae in pure RO, 1/2 RO-tap, GH=4 KH=0

Experiment 3:Treat with Easylife Carbo at 1x, 2x, 5x, 10x

The problems with this experiment incluse the lack of organic substances from a normal aquarium. Lack of water movement... accepting donations to buy mixers or agitators.

Do you see any problems or confounders ?
What other tests would you like to add ?
 
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#2 ·
Very interesting Mihai. I'd also try to test the samples in very high nutrients rich water (like double EI?) and in water with saturated Co2. I'd also try to put the samples in water rich of organic waste to see if it grow more than in clean water. Very curious to see the results!
 
#4 ·
Thank you very much for your support and feedback. Keep it coming. :grin2:
I'd also try to test the samples in very high nutrients rich water (like double EI?) and in water with saturated Co2. I'd also try to put the samples in water rich of organic waste to see if it grow more than in clean water.
Thank you for your great ideas @fablau. The one with 2x EI water is easily done, I will try to start a sample. Saturated CO2 and water rich in organic waste are a bit harder to do and measure. Maybe after this round of experiments ends I will have some good / easy ( read lazy) solutions to conduct the experiments.

Mihai,
Unfortunately, I was never successful, even if I moved them within the same aquarium they came from. They just melted away.
Hmm, interesting input @g4search. I agree ideally I would expose established colonies. Maybe I can put some chopsticks in the aquarium and wait for the algae to colonize them. Afterwards, I can cut it in smaller pieces and expose the established colonies. What do you think about this ?
I tried to have at least one complete unit in one treatment. A mature “unit” can release spores, but it is not realistic to wait for them to grow (time and glass). Also, this is one reason I have the control sample. It has tap water with EI. If the algae there is still alive and the one in the treatment bottle dies, then we can suggest that the treatment is responsible for the death.

Today I exposed one sample to light after a 3 day lights-out. It is still alive. I will come back with a post with photographs showing the interim results.
 
#3 ·
Mihai,
it sounds good what you are planning. However, I just want you to know that I have tried several times to "transplant" BBA and make them grow in different containers. Unfortunately, I was never successful, even if I moved them within the same aquarium they came from. They just melted away. I have the feeling BBA has to grow from spores, and once they are in the growth-phase they can't be "uprooted" and grown elsewhere.
 
#11 ·
I used 5x dose of Excel for about two months in a tank with fish (lots of young bettas) and plants trying to control BGA outbreak. Everything was fine - both fish and plants (mostly crypts) did well. BGA stopped spreading from day one of treatment and slowly started to retreat but didn't disappear completely even after about 2 months of such dosing.
 
#13 ·
@dukydaf, Mihai, I was under the impression that you actually wanted to "grow" BBA, so you could eventually treat the growing BBA to figure out how to prevent further growth (or perhaps stop sporulation). This would be the most rational way for you to actually control BBA.
My interpretation of your current experiments is that you are treating various trans-located BBA specimens to see which one gets more discolored? (as indication of DEAD specimen?) I'm sure that you can keep your "control" specimen for 3-4 weeks without notable changes, but the REAL question is: is it still truly alive? (look at it as cut flowers. You can keep them in a vase for one or two weeks, but eventually you have to throw them out because they are dead)

You know, I think just about everybody on this site here know how to kill BBA ( e.g. spot treatment w glut or H2O2 [or hypoclorite]). I am sure there are many more "methods" to kill BBA. It is actually not very difficult! Prevention on the other hand, is much much harder. ( Theoretically, if you could keep BBA spores out of your tank, you would NEVER have a BBA problem. That seems to be unrealistic however.)

So the only other route then is to control its growth. There are certainly many factors that have to be considered. I my opinion, LIGHT using the proper wavelength spectrum is the most important factor! At this stage, what I can tell you is that light at wavelengths around 410 nm (at what intensity?) are absolutely necessary for BBA to grow.

BTW, with respect to your pictures here, they are excellent! I am sure that nobody here would ever question whether they are doctored.
 
#16 · (Edited)
Hello fellow algae growers,

Though it was time for a weekly update. In short: easy carbo 1x kills BBA detached or on leaf. Differences in water type is inconclusive. Putting BBA in the dark for one week does nothing. Experiments with BBA and organic matter were inconclusive (yet).

Experiment 1: Day 14

No difference in the way algae look. But there is some string algae developing in the RO water and 1/2 tap.


Experiment 2: Day 14 - only control and 1x shown



8 days after stopping the experiment, the 1x sample is still dead white. Control still going strong. All other samples looked like 1x.


Experiment 3: Day 14


Nothing different in the algae held in the darkness.

Experiment 4: Day 7

Experiment 4: Description
Taking into consideration the feedback received I tried to experiment on algae on leaves. I tried to expose the algae to organic material.

Leaf extract = 5 leaves of H. pinnatifida grinded in 10ml water.(not a scientific way to measure but should still release a lot of what is in the leaf)
Crushed solution= 2ml of leaf extract in 8ml water (w/ macros, see first post)
crushed 1/2 = 1ml of leaf extract+9ml water

I also tried the CO2 experiment once again (1x), in water with no organics and in water with organics.

Experiment 4: Results

It is a little difficult to see in the photos from Experiment 4. In the Easycarbo 1x treated bottles (marked with CO2), both the freestanding algae and the algae on the leaves was dead. In crushed 1/2 and crushed samples with BBA on Althernathera, the algae is still there, not necessarily growing but healthy. In the "crushed" sample with no leaf, the detached colony of BBA is dead and white.

In sample(not shown) with leaf extract and BBA on H. pinnatifida leaf, the BBA turned white and now only low patches of BBA are visible (looks like GDA but with the color of BBA).

Maybe H. pinnatifida has some algae killing toxin in the cells that was in high enough concentration. I will try with snail poop and fish food next, see if I manage to get some rapid propagation. The problem that I see is that green algae also develop in these conditions and might outcompete the BBA.
 
#22 · (Edited)
Some short news first:

The chopstick in the aquarium is beginning to be colonized by BBA, many small colonies at the moment. Took about 2 weeks. I will keep the stick in there so that they continue to grow.

The control sample for fish food was severely contaminated and I need to restart the experiment.

The sample I kept in the dark for 23 days looks fine, like nothing happened...

I wonder what you can do to show me that the BBA in your "control" experiment is actually still alive?
(Just because the color didn't change, does not mean the organism is still viable)
Looking forward to your next set of experiments!
Thank you @g4search for closely following these experiments and being skeptical about the results. What do you mean by viable ? If anybody sees any problems with my reasoning or methods please let me know. Better to have doubts in the right places than be ignorant. Ah, what is alive and what is dead ? Very hard to define life if you think about it, but I will try to do so with BBA.

As previously stated, to undoubtedly prove that something is alive we need establish that it is capable of metabolic activity and multiplication. For algae that would mean photosynthesis during light hours, maybe measured as oxygen production, and spore production. I don't have the equipment necessary at home to measure that, so I use surrogate endpoints. Would be nice to have a lab though :D. Multiplication was not yet observed and may come from contamination with spores already available.

Example of surrogate endpoint: long term integrity of the cell and cell organelles under normal conditions as a good surrogate for a live cell. For me this is a good surrogate. Very few organisms manage to maintain their structural integrity for a long time after death (again, normal conditions). Additionally, pigments like chlorophyll would be one of the first to degrade if the organism has died so if we observe no chlorosis (transparent/white leaves) we can assume there is still metabolic activity.

Another way of thinking, less scientific but practical nevertheless is .... does it look like a dead BBA ? Does it act like a dead BBA ? Do you have any reason to assume it is a dead BBA ? If no, assume it is not dead.

I didn't specialize in algae, so maybe somebody with more information on algae physiology can comment. Do we have any reason to think that the moment a BBA colony is detached it dies ?

Now your turn :D Can you prove this BBA colony is dead ?


another interesting experiment to try is to see resistance of BBA to toxicity levels of CSM+B. Maybe at different concentrations?
Thank you for the link to thread, didn't have time to read all of it. I like the idea behind the bacterial population as an intermediary, but of little relevance to our aquariums. It is an interesting theory, though I highly doubt BGA would be affected by CSM levels that are not toxic to plants and invertebrates. I don't have access to CSM+B in the EU but can test it with another trace mix such as JBL Ferropol.

More coming soon.
 
#29 · (Edited)
Other acids will require the reduction of carbonate concentration to reach a lower pH endpoint. CO2 will affect H+ concentration without (significantly) affecting CO3 concentration.

With these samples it should become clear whether it's H+ concentration, CO2 concentration, CO3 concentration or some combination that has the greatest effect on BBA.


  • High H+, low CO3
  • Neutral H+/OH-, low CO3
  • High OH-, high CO3
  • High H+ (high CO2), low CO3
  • High H+ (very high CO2), high CO3
  • Neutral H+/OH- (high CO2), high CO3
  • High OH- (no CO2), high CO3

The first three samples would have CO2 concentration at equilibrium with the atmosphere. Lots of surface agitation will help to ensure consistent CO2 concentration, and minimize any effects on CO2 concentration from the transformation of CO3 to HCO3 to H2CO3 <> CO2 + H2O. Ideally the acid injection would be maintained with CO3 injection to maintain a steady concentration of both, however, given the (overall) sample size, a reasonable conclusion can probably be obtained even with samples varying to some degree around a base concentration. If you're after ideas, I would setup an acid solution and bicarbonate solution with some form of consistent (think minutes, not days) dosing. A balance of dosing should be found to target pH endpoint, where monitoring of KH and pH will inform of correction needed.

If KH low and pH drops > increase HCO3 injection.
If KH correct and pH drops > reduce acid injection.


The next three samples would be significantly easier to control since CO2 injection is easy to maintain consistently, and doesn't affect CO3.

The last sample would be extremely difficult to control (read: I don't know how), since I know of no way to remove CO2 from water with stability over time. A large plant mass could probably easily reduce CO2 concentration, however the concentration would be variable with variable light. edit: Actually this variability in concentration probably isn't such a bad thing since it will more accurately represent real life conditions.
 
#31 ·
A form of peat extract would be better imo.

There is some consensus that it is the variability of things that promotes algae, and hence any experiments to determine the effects of a thing need to control variability. Otherwise we don't know if it was the concentration of the thing that had the effect, or the variability of the thing, or some combination.
 
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