What are the root causes of Cyano?

[taricha]

Brilliant experiment with sodium nitrate! How long would you say it took for the cyanobacteria to show up? I don’t want to cut my experiment short.

By 1-2 weeks the colony was well established. Once established it added that much tissue every couple of days.

 

[Dan_P]

Often cyano is the only thing found growing in a dino mat, and dinos in cyano mats. Whose mat is it anyway? RumMnky's pic isn't an aberration. They feel more like accomplices to me than competitors.

Funny you mention that nutrient strategy, I wanted to see what would happen if I put my normal sodium nitrate dose pellets in one spot instead of dissolved into flow. So i've been dropping them in a beaker in my sump, where water flows from beaker into skimmer chamber.
Guess what grows on the beaker?
Fat blobs and sheets of spirulina.

I want to estimate NO3 in your flow. What would you estimate is the water flow from the beaker? How long does the sodium nitrate last in the beaker? How much sodium nitrate is in the beaker?

 

[Dan_P]

By 1-2 weeks the colony was well established. Once established it added that much tissue every couple of days.

Not long at all. With observations using a microscope, I should see the buildup sooner. Thanks

 

[MabuyaQ]

It does make you wonder why this pest is so difficult to grow. Do we know of dinoflagellates preventing cyanobacteria growth?

My next attempt will be to use nutrients at the level used to cultivate bacteria, 100’s of ppm nitrate and 20 ppm phosphate. What cyanobacteria wouldn’t like that?

Both are capable of producing allelopathic substances, but to what extend these can supress eachother or even stimulate eachother I really don't know and is probably species specific as well. We just can't rule out that this species of dino is able to surpress that species of cyano under favourable conditions for that dino species, which may very well be the same or even broader than that of this cyano species as well. Making it impossible for a cyano outbreak to happen in your experiment unless you take this dino out of the equation.

Start with sterile substrate, sterile seawater, isolate you cyano strain and restart the tests. If you can trigger a cyano outbreak in the absence of this dino, you can then also test what happens if you reintroduce it (in great numbers so keep cultering them as well) besides testing for different cyano outbreak triggers.

 

[Dan_P]

Both are capable of producing allelopathic substances, but to what extend these can supress eachother or even stimulate eachother I really don't know and is probably species specific as well. We just can't rule out that this species of dino is able to surpress that species of cyano under favourable conditions for that dino species, which may very well be the same or even broader than that of this cyano species as well. Making it impossible for a cyano outbreak to happen in your experiment unless you take this dino out of the equation.

Start with sterile substrate, sterile seawater, isolate you cyano strain and restart the tests. If you can trigger a cyano outbreak in the absence of this dino, you can then also test what happens if you reintroduce it (in great numbers so keep cultering them as well) besides testing for different cyano outbreak triggers.

Good suggestions. If “nothing” continues to happen, I will have to develop a more rigorous technique to grow cyanobacteria.

 

[Dan_P]

any pictures possible?

Here is a close up of one of the scattered and very long pink filaments living amongst the sand grains in Cyano Culture 1 (now dismantled). Is it possible that I am seeing groupings of four cells in the trichome or is that an artifact?

 

[taricha]

I want to estimate NO3 in your flow. What would you estimate is the water flow from the beaker? How long does the sodium nitrate last in the beaker? How much sodium nitrate is in the beaker?

1g of NaNO3, dissolved in (originally empty) 500mL beaker in 5 min.
In that time, 1.5L flowed in beaker.
So a very short bust of ~400ppm NO3 then a rapid drop down to ~20ppm as it exchanges water with a smallish chamber then over the next hour or two that small chamber reaches equilibrium with the tank in the 5ppm ballpark.

 

[Frogger]

Hi Dan

Not sure if this would be of any help but I had a cyano bloom recently (not spirolina) after a large fish died (couldn't find) which spiked my nitrates up to above 40ppm for over a month. I used GFO to bring it back down. When it came down it came down rapidly to undetectable amounts. It was at this point that the cyano bloomed. It covered all rocks and gravel as it is effective at fixing nitrates out of rock when there is none in the water column. Maybe either artificially raise your nitrates up for an extended period, or get some rocks from a tank where there is high nitrates. Once the rock has absorbed the nitrates use GFO or other nitrate removing methods to quickly lower the nitrates to below detectable amounts. This should get a good batch of cyano growing.

 

[Dan_P]

1g of NaNO3, dissolved in (originally empty) 500mL beaker in 5 min.
In that time, 1.5L flowed in beaker.
So a very short bust of ~400ppm NO3 then a rapid drop down to ~20ppm as it exchanges water with a smallish chamber then over the next hour or two that small chamber reaches equilibrium with the tank in the 5ppm ballpark.

1g of NaNO3, dissolved in (originally empty) 500mL beaker in 5 min.
In that time, 1.5L flowed in beaker.
So a very short bust of ~400ppm NO3 then a rapid drop down to ~20ppm as it exchanges water with a smallish chamber then over the next hour or two that small chamber reaches equilibrium with the tank in the 5ppm ballpark.

If this were repeatable with a control (beaker, no sodium nitrate dose), we would spend some energy discussing why short burts of nitrate stimulate cyanobacteria growth. With the level of data that we have, the explanation “a coincidence” is a viable one.

But in a nod to the schoolof thought of “high nutrient levels are important”, I started Cyano Culture 2 with a small mat of cyanobacteria and very likely some GHA debris, and a culture medium of aquarium water spiked with NO3 (281 ppm), PO4 (5 ppm) and iron (II) gluconate (0.2 ppm).

 

[Dan_P]

Hi Dan

Not sure if this would be of any help but I had a cyano bloom recently (not spirolina) after a large fish died (couldn't find) which spiked my nitrates up to above 40ppm for over a month. I used GFO to bring it back down. When it came down it came down rapidly to undetectable amounts. It was at this point that the cyano bloomed. It covered all rocks and gravel as it is effective at fixing nitrates out of rock when there is none in the water column. Maybe either artificially raise your nitrates up for an extended period, or get some rocks from a tank where there is high nitrates. Once the rock has absorbed the nitrates use GFO or other nitrate removing methods to quickly lower the nitrates to below detectable amounts. This should get a good batch of cyano growing.

Thanks for the detailed recap. I have seen cyanobacteria cover sand, rocks, macro algae, PVC, and glass. Did the phosphate level increase as well?

Cyano Culture 2 just started with 1500 mL of aquarium water spiked with NO3 (281 ppm), PO4 (5 ppm) and iron (II) gluconate (0.2 ppm) in which I added a piece of a cyanobacteria mat I removed from the sump. Since I can’t seem to induce a cyanobacteria mat to form, I need to know whether I can even keep one going.

 

[Frogger]

hanks for the detailed recap. I have seen cyanobacteria cover sand, rocks, macro algae, PVC, and glass. Did the phosphate level increase as well?

Cyano Culture 2 just started with 1500 mL of aquarium water spiked with NO3 (281 ppm), PO4 (5 ppm) and iron (II) gluconate (0.2 ppm) in which I added a piece of a cyanobacteria mat I removed from the sump. Since I can’t seem to induce a cyanobacteria mat to form, I need to know whether I can even keep one going.

My bad. I just went back over my reef log, I was brain dead earlier today. My phosphates shot up after the tang died my nitrates were pretty stable at around 1.0 ppm.
Normally my phosphates are naturally low in my tank it usually hovers around .01ppm of PO4 and I have to add KNO3 to keep my nitrates above zero. After the tang died my phosphates shot up to over .12ppm or 40 on the Hanna low range phosphorus meter. After my tang died hair algae growth went nuts with the high Phosphate and no tang. I stopped adding KNO3 and ran GFO. Once the phosphate dropped down to less than .03 my nitrates dropped to zero (Red Sea Pro test kit) and cyano started growing. Once it started I couldn't stop it. It took me over 6 months for my tank to stabilize and stop the cyano from growing.

One other thing that seemed to give the cyano a good kick start was I tried dosing vibrant to stop the hair algae growth when the phosphates were high.

 

[bigdrew]

It seems like I start getting some cyano when my nitrates and phos are very low. I start feeding a bit more and it slowly goes away.

This is my situation right now. I shut off my skimmer, started feeding a little more and have barely upped my Nitrate from 0 to maybe 1 or so. It has already had an impact on the cyano.

Also, always remember to keep that TDS meter handy. Just when you think the RO or RODI filter is brand new and couldn’t be putting out 1 or higher TDS readings, think again. Worth checking Every. Single. Time.

 

[Dan_P]

My bad. I just went back over my reef log, I was brain dead earlier today. My phosphates shot up after the tang died my nitrates were pretty stable at around 1.0 ppm.
Normally my phosphates are naturally low in my tank it usually hovers around .01ppm of PO4 and I have to add KNO3 to keep my nitrates above zero. After the tang died my phosphates shot up to over .12ppm or 40 on the Hanna low range phosphorus meter. After my tang died hair algae growth went nuts with the high Phosphate and no tang. I stopped adding KNO3 and ran GFO. Once the phosphate dropped down to less than .03 my nitrates dropped to zero (Red Sea Pro test kit) and cyano started growing. Once it started I couldn't stop it. It took me over 6 months for my tank to stabilize and stop the cyano from growing.

One other thing that seemed to give the cyano a good kick start was I tried dosing vibrant to stop the hair algae growth when the phosphates were high.

When hair algae or Caulerpa appear to be growing weakly in my system’s sump, cyanobacteria mats cover them (Dinoflagellates too). Another semi regular event in the display tank is Caulerpa becoming covered with sand and then the sand being covered with cyanobacteria. I think @brandon429 would not be surprised to hear about the latter. I am wondering if the overpopulation of starving algae was the real cause of the cyanobacteria growth.

I am still being humiliated by the lack of cyanobacteria growth and might have to supply some organic matter to my culture medium.

 

[Dan_P]

I proved that I could nurture a piece of cyanobacteria mat to grow and spread (Cyano Culture 2). It was a small piece that I was able to adhere to a piece of sea glass glass. It was submersed in aquarium water spiked with nitrate and phosphate. And I think observing growth rates of little bits of existing mats adhered to glass as I vary nutrient types and concentrations will be my first step to understanding cyanobacteria mat formation.

Pictured below is my latest attempt to adhere a small piece of mat to a microscope slide cover slip (very thin glass about 2 cm x 2 cm). This particular mat consists of more than one organism. Over a twenty four period, it appears that the green cyanobacteria is moving away (not necessarily growing) from the darker core. If you look closely, you can see the mat is surrounded by a green haze on the cover slip, presumably migrating Spirulina. I still need to dissect this glob to see whats happening.

 

[taricha]

. This particular mat consists of more than one organism. Over a twenty four period, it appears that the green cyanobacteria is moving away (not necessarily growing) from the darker core. If you look closely, you can see the mat is surrounded by a green haze on the cover slip, presumably migrating Spirulina. I still need to dissect this glob to see whats happening.

Oh dear. Now you've initiated the Great Cyano Schism. What have you done?!?

 

[Dan_P]

Oh dear. Now you've initiated the Great Cyano Schism. What have you done?!?

This might turn out to be a microbial Game of Thrones. Stay tuned.

 

[Dan_P]

OK, here are the results of my first cyanobacteria nutrition study. I tested the nutrient treatments in triplicate. The treatments were NO3 200 vs 20 ppm and PO4 4 v 0.4 ppm. I measured the amount of chlorophyll extracted from each culture as a way to estimate biomass. The bar height in the graph below corresponds to the amount of chlorophyll present in the culture. The variation (error bar size) for the 200 ppm NO3/0.4 ppm PO4 treatment was large, making it impossible to say that its chlorophyll level was different from the 200 ppm/4 ppm treatment.

It appears that nitrate level had a significant effect for the range studied but PO4 didn’t. I guess that PO4 had no affect on growth only because the low dose was more than the bacteria needed. Would be interested in your thoughts.

In my second nutrition study, I am looking at ammonia and phosphate.

 

[MnFish1]

OK, here are the results of my first cyanobacteria nutrition study. I tested the nutrient treatments in triplicate. The treatments were NO3 200 vs 20 ppm and PO4 4 v 0.4 ppm. I measured the amount of chlorophyll extracted from each culture as a way to estimate biomass. The bar height in the graph below corresponds to the amount of chlorophyll present in the culture. The variation (error bar size) for the 200 ppm NO3/0.4 ppm PO4 treatment was large, making it impossible to say that its chlorophyll level was different from the 200 ppm/4 ppm treatment.

It appears that nitrate level had a significant effect for the range studied but PO4 didn’t. I guess that PO4 had no affect on growth only because the low dose was more than the bacteria needed. Would be interested in your thoughts.

In my second nutrition study, I am looking at ammonia and phosphate.

Curious - how did you isolate 'cyanobacteria' as the only chlorophyll containing organism in your experiment?
Second - Without doing the statistics - I would ask - is there a statistically significant difference between Bar 2 (200/4) and Bar 4 (20/.4) - if not - then you cant really say there is any difference between any of them.
Third is there a significant difference between Bar 2 and 3 - if not - its hard to make the case that there is any difference.

Having said that - I'm assuming you're comparing Bar 1 (200/4) to Bar 3(20/4) so suggest that there is an effect of nitrate. Did you see the same significant between Bar 2 and Bar 4? (if not - it could be that the standard of error in Bar 2 - is the error - as compared to bar 2...?

If I were you I would (rather than doing another study with ammonia and PO4 - would instead repeat the first one - to see why the results were so disparate.


BTW - totally appreciate your efforts here - its quite interesting. It would be really nice to see the actually p values/ whatever statistics you used to analyze differences between the bars.

 

[Dan_P]

Curious - how did you isolate 'cyanobacteria' as the only chlorophyll containing organism in your experiment?
Second - Without doing the statistics - I would ask - is there a statistically significant difference between Bar 2 (200/4) and Bar 4 (20/.4) - if not - then you cant really say there is any difference between any of them.
Third is there a significant difference between Bar 2 and 3 - if not - its hard to make the case that there is any difference.

Having said that - I'm assuming you're comparing Bar 1 (200/4) to Bar 3(20/4) so suggest that there is an effect of nitrate. Did you see the same significant between Bar 2 and Bar 4? (if not - it could be that the standard of error in Bar 2 - is the error - as compared to bar 2...?

If I were you I would (rather than doing another study with ammonia and PO4 - would instead repeat the first one - to see why the results were so disparate.


BTW - totally appreciate your efforts here - its quite interesting. It would be really nice to see the actually p values/ whatever statistics you used to analyze differences between the bars.

All valid points. My statical analysis was rather simplistic. Let me see if I can upgrade the analysis. And repetition is definitely on the books.

The Spirulina cultures were not pure, so, kudos for the insightful question. Why I think the chlorophyll was mostly of cyanobacteria origin is the lack of significant dinoflagellate pigment, the only other significant photosynthetic biomass. Admittedly, this not a strong defense

 

[Dan_P]

Curious - how did you isolate 'cyanobacteria' as the only chlorophyll containing organism in your experiment?
Second - Without doing the statistics - I would ask - is there a statistically significant difference between Bar 2 (200/4) and Bar 4 (20/.4) - if not - then you cant really say there is any difference between any of them.
Third is there a significant difference between Bar 2 and 3 - if not - its hard to make the case that there is any difference.

Having said that - I'm assuming you're comparing Bar 1 (200/4) to Bar 3(20/4) so suggest that there is an effect of nitrate. Did you see the same significant between Bar 2 and Bar 4? (if not - it could be that the standard of error in Bar 2 - is the error - as compared to bar 2...?

If I were you I would (rather than doing another study with ammonia and PO4 - would instead repeat the first one - to see why the results were so disparate.


BTW - totally appreciate your efforts here - its quite interesting. It would be really nice to see the actually p values/ whatever statistics you used to analyze differences between the bars.

Here are the stats for the model

Abs @ 656 = NO3 + PO4 + Constant.

The interaction term was not statistically significant. R2 for the model was 0.6.

What do you think?