What are the root causes of Cyano?

[MnFish1]

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?

I dont know what intercept is. I would just take the mean and SD and do a t-test. (is that what you did)? Again - its unclear which tubes you're comparing?

 

[Dan_P]

I dont know what intercept is. I would just take the mean and SD and do a t-test. (is that what you did)? Again - its unclear which tubes you're comparing?

Relook at the bar chart X-axis. The treatment is under each bar. “200/4” means 200 ppm NO3 and 4 ppm PO4. Also note the error bar on each bar. That represents 1 standard deviation.

The new table displaying NO3, PO4 and Intercept are the factors of the linear equation that describe the data. A P-value for each factor is the statistic that describes the likelihood that the level of its influence happens by chance. The NO3 value indicates that its level of influence happens by chance about 1 in a 1000 where as the level of PO4 influence happens by chance 39 in a hundred. If we use 5 in a 100 as a cut off for statistical significance, we would say that in my experiment, PO4 had no significant effect while NO3 had a very significant effect. The R^2 value describes how well the model describes the variation in the data.

Clearer now?

 

[taricha]

Oh, one Q. Is it correct to say the cyano mats grew under all the treatments, but much moreso under the high NO3 condition? Or is it not clear if the low(er) NO3 samples had much cyano growth?

 

[Dan_P]

Oh, one Q. Is it correct to say the cyano mats grew under all the treatments, but much moreso under the high NO3 condition? Or is it not clear if the low(er) NO3 samples had much cyano growth?

Based on the amount of chlorophyll extracted, “yes” to your first question.

 

[MnFish1]

Relook at the bar chart X-axis. The treatment is under each bar. “200/4” means 200 ppm NO3 and 4 ppm PO4. Also note the error bar on each bar. That represents 1 standard deviation.

The new table displaying NO3, PO4 and Intercept are the factors of the linear equation that describe the data. A P-value for each factor is the statistic that describes the likelihood that the level of its influence happens by chance. The NO3 value indicates that its level of influence happens by chance about 1 in a 1000 where as the level of PO4 influence happens by chance 39 in a hundred. If we use 5 in a 100 as a cut off for statistical significance, we would say that in my experiment, PO4 had no significant effect while NO3 had a very significant effect. The R^2 value describes how well the model describes the variation in the data.

Clearer now?

Kind of - but I don't agree with your conclusions. Firstly - I would think that you would need to test many more than 2 ratios to show what you're trying to say. I.e. 2 measurements doesn't really show a dose response curve. In other words - what is the basis for choosing the ratios you did? Seems like you also need a control (i.e. nitrate alone - and Phosphate alone) as well as one with neither nitrate or phosphate.

The error is so high on the second sample (as you said) Its impossible to say IMHO - that PO4 has no effect at high levels of nitrate.

But its interesting to look at. How are you sure that you have a pure culture of cyanobacteria - as compared to cyanobacteria + some other algae etc? This would also confound the results. Did you sample at different levels of light/time of day?

BTW I really appreciate your efforts - most people wouldn't take the time to do the experiment. One other question - what was your hypothesis - or did you just randomly choose concentrations of NO3 and PO4?

 

[MnFish1]

Based on the amount of chlorophyll extracted, “yes” to your first question.

What was the answer to the second question?

 

[Dan_P]

What was the answer to the second question?

T

What was the answer to the second question?

The second question was the opposite of question one, so, the answer is “no” because I could tell and got great stats on the nitrate effect.

 

[MnFish1]

Relook at the bar chart X-axis. The treatment is under each bar. “200/4” means 200 ppm NO3 and 4 ppm PO4. Also note the error bar on each bar. That represents 1 standard deviation.

The new table displaying NO3, PO4 and Intercept are the factors of the linear equation that describe the data. A P-value for each factor is the statistic that describes the likelihood that the level of its influence happens by chance. The NO3 value indicates that its level of influence happens by chance about 1 in a 1000 where as the level of PO4 influence happens by chance 39 in a hundred. If we use 5 in a 100 as a cut off for statistical significance, we would say that in my experiment, PO4 had no significant effect while NO3 had a very significant effect. The R^2 value describes how well the model describes the variation in the data.

Clearer now?

PS - here is an interesting article on p Values. I think it has a bearing on your results. https://www.amstat.org//asa/files/pdfs/P-ValueStatement.pdf

 

[Dan_P]

PS - here is an interesting article on p Values. I think it has a bearing on your results. https://www.amstat.org//asa/files/pdfs/P-ValueStatement.pdf

Thanks! Great article.

 

[Dan_P]

Kind of - but I don't agree with your conclusions. Firstly - I would think that you would need to test many more than 2 ratios to show what you're trying to say. I.e. 2 measurements doesn't really show a dose response curve. In other words - what is the basis for choosing the ratios you did? Seems like you also need a control (i.e. nitrate alone - and Phosphate alone) as well as one with neither nitrate or phosphate.

The error is so high on the second sample (as you said) Its impossible to say IMHO - that PO4 has no effect at high levels of nitrate.

But its interesting to look at. How are you sure that you have a pure culture of cyanobacteria - as compared to cyanobacteria + some other algae etc? This would also confound the results. Did you sample at different levels of light/time of day?

BTW I really appreciate your efforts - most people wouldn't take the time to do the experiment. One other question - what was your hypothesis - or did you just randomly choose concentrations of NO3 and PO4?

All good points. Many will be addressed with further experimentation. Thanks,
Dan

 

[Dan_P]

I recently completed two more nutrient experiments on the Spirulina from my aquarium. Each experiment looked at low and high PO4 levels and low and high NH3 levels (design of experiments). The first experiment tested NH3 below 0.1 ppm N-NH3 while the seconded looked at NH3 levels above 0.1 ppm N-NH3. The amount of chlorophyll A extracted from the cyanobacteria was used as an indicator for growth. Results from both experiments are plotted on the same graph. The blue line are the high PO4 results, the lower yellow line the low PO4 condition. The dashed line connects the results of the two experiments.

In general, when more nitrogen is available, more growth occurs (rising slope of the line). An interesting result occurs at the low PO4 level (yellow line), where the increased growth from increasing ammonia nitrogen flattens out near 0.1 N-NH3, (1.7 ppm NH3). The cyanobacteria cannot utilize the extra ammonia because it is PO4 limited. Given enough PO4, growth continues to increase with increasing nitrogen (blue line).

I still have not found the “trigger” for cyanobacteria mat formation. My experiments are generating thin films, suggesting to me that my experiment conditions although nutrient rich are not sufficient to create those aggravating mats. I begin to doubt whether the nitrate and phosphate levels in aquarium water are the sole cause for cyanobacteria mat formation.

The search continues.

 

[Dan_P]

Just a quick update. I am currently involved with Cyano Nutrition Study #6. 24 Spirulina cultures with varying amounts of nitrate, phosphate and skimmate as a source of DOC. @Rick Mathew and @taricha are both advising me, reviewing data and developing ideas.

While I can grow thin films of cyanobacteria, none resemble the “luxurious” growth seen in our aquaria and refugia. The root causes or triggers we have discussed in this thread are beneficial to growth but do not produce aggressive growth or thick mats. There seems to be more to mat formation than a certain level of nitrate, phosphate, iron or DOC in the aquarium.

Our small offline “cyano club” has come to consider the possibility that a cyanobacteria mats may be a sort of collaborative effort of aerobic heterotrophic bacteria and cyanobacteria. A cyanobacteria may not be able to form a thick film or mat without the right kind of aerobic heterotrophic bacteria partner. Nutrients play a supporting role in mat formation, but without the partnership, no amount of nutrient will stimulate mat formation. We are currently discussing ways to test this idea or to use this hypothesis to grow heavy mats in a “test tube”. One outcome of this discussion was to start add commercially available bottled bacteria to cyano cultures. Another was to start adding various types of dissolved organic carbon. As you might guess, the number of experiments to conduct is ballooning.

 

[Belgian Anthias]

The route cause of cyano outbrakes in a closed environment is lack op competition. Cyano where first, to have a change other photo-autotrophs and heterotrops had to grow faster.( CMF De Haes)

Most marine cyano are not bentic and do not form heterocysts> A lot live in symbiosis with diatoms and other organisms. Also in aquaria cyano are everywhere and they should be. A lot of them will be able to form heterocysts.
They play an important role in the coral holobiont and in and on most lower animals.
There function in microbial matts is what most reefkeepers dislike about them. They play a main role in about any ecosystem.

Cyanobacteria occupy a unique position within the group of nitrogen-fixing organisms. Cyanobacteria are the only oxygen-producing photosynthetic organisms that are capable of fixing nitrogen gas in combination with carbon dioxide fixation. It is known there growth can be controled by lightintensity (spectrum+energy) as they stop growing at high light intensity and will die after a period of exposure. Most photo-autotrophs are able to protect themself against high light intensity by synthesising the xanthofyl cyclus, cyano are not able to do so.

Battling them by nutrient limitation is possible but not without effecting all other live in the closed environment of an aquarium.
Addition of carbohydrates will increase heterotrophic growth and will outcompete cyano in the watercolumn.
Cyano, part of a microbial matt, growth will be influenced by increased oxygen consumption on the outer layers of the matt and changing light conditions. In such conditions there function is providing oxygen to its companions in the matt. Such a matt may be selfsuporting for a long period, even with no nutrients in the watercolumn. Cyano are part of the best growing photo-autotrophs in low light conditions.



The cyano site: http://www-cyanosite.bio.purdue.edu/images/images.html

 

[Belgian Anthias]

The route cause of cyano outbrakes in a closed environment is lack op competition. Cyano where first, to have a change other photo-autotrophs and heterotrops had to grow faster.( CMF De Haes)

Most marine cyano are not bentic and do not form heterocysts> A lot live in symbiosis with diatoms and other organisms. Also in aquaria cyano are everywhere and they should be. A lot of them will be able to form heterocysts.
They play an important role in the coral holobiont and in and on most lower animals.
There function in microbial matts is what most reefkeepers dislike about them. They play a main role in about any ecosystem.

Cyanobacteria occupy a unique position within the group of nitrogen-fixing organisms. Cyanobacteria are the only oxygen-producing photosynthetic organisms that are capable of fixing nitrogen gas in combination with carbon dioxide fixation. It is known there growth can be controled by lightintensity (spectrum+energy) as they stop growing at high light intensity and will die after a period of exposure.
Most photo-autotrophs are able to protect themself against high light intensity by synthesising the xanthofyl cyclus, cyano are not able to do so.

Battling them by nutrient limitation is possible but not without effecting all other live in the closed environment of an aquarium.
Addition of carbohydrates will increase heterotrophic growth and will outcompete cyano in the watercolumn.
Cyano, part of a microbial matt, growth will be influenced by increased oxygen consumption on the outer layers of the matt and changing light conditions. In such conditions there function is providing oxygen to its companions in the matt. Such a matt may be selfsuporting for a long period, even with no nutrients in the watercolumn. Cyano are part of the best growing photo-autotrophs in low light conditions.

 

[taricha]

Our small offline “cyano club” has come to consider the possibility that a cyanobacteria mats may be a sort of collaborative effort of aerobic heterotrophic bacteria and cyanobacteria. A cyanobacteria may not be able to form a thick film or mat without the right kind of aerobic heterotrophic bacteria partner. Nutrients play a supporting role in mat formation, but without the partnership, no amount of nutrient will stimulate mat formation. We are currently discussing ways to test this idea or to use this hypothesis to grow heavy mats in a “test tube”. One outcome of this discussion was to start add commercially available bottled bacteria to cyano cultures.

Here's some food for thought for those following along...
Test tubes on day 16.
Normal light vs Cyano Fluorescence - (red cyano pigment glows red under green LED) - for 4 different treatments - with duplicates of each.
1. Control
2. Vibrant
3. Chaeto only
4. Vinegar

[experiment notes: All tubes got seeded with a slurry of cyano cells, the control got nothing else. The other 3 treatments got the same 0.06g of frozen/thawed chaetomorpha as a dying algae food for the cyano. Vinegar tretment got a drop of vinegar on days 1, 4, 7. Vibrant treatment got a drop of vibrant on days 1,4,7.]

Control and Chaeto treatments: as expected, some nutrients from water allowed the small number of cyano cells to grow a little bit. Dead leaking chaetomorpha grew somewhat more than the control.

It seems Vibrant either directly helps cyano or support cyano friendly bacteria indirectly?
Vinegar grew a bacterial biofilm on the water's surface, but it apparently was not the kind of bacteria that plays well with cyano, as it suppressed cyano growth and had the least cyano of any treatment.

what will other bacterial bottle products show? hmm....

 

[Dan_P]

Here's some food for thought for those following along...
Test tubes on day 16.
Normal light vs Cyano Fluorescence - (red cyano pigment glows red under green LED) - for 4 different treatments - with duplicates of each.
1. Control
2. Vibrant
3. Chaeto only
4. Vinegar

[experiment notes: All tubes got seeded with a slurry of cyano cells, the control got nothing else. The other 3 treatments got the same 0.06g of frozen/thawed chaetomorpha as a dying algae food for the cyano. Vinegar tretment got a drop of vinegar on days 1, 4, 7. Vibrant treatment got a drop of vibrant on days 1,4,7.]

Control and Chaeto treatments: as expected, some nutrients from water allowed the small number of cyano cells to grow a little bit. Dead leaking chaetomorpha grew somewhat more than the control.

It seems Vibrant either directly helps cyano or support cyano friendly bacteria indirectly?
Vinegar grew a bacterial biofilm on the water's surface, but it apparently was not the kind of bacteria that plays well with cyano, as it suppressed cyano growth and had the least cyano of any treatment.

what will other bacterial bottle products show? hmm....

Here I am beavering away looking for nutrients to grow luxurious cyanobacteria mats and someone is selling a “probiotic” that will grow strong and healthy cyanobacteria! I am going to get me a bottle -)

Pretty cool putting your knowledge on visible light to practical use.

Did you get around to looking at the cyano under the microscope? I am curious about what types of organisms live in the mat. I find it varies depending on the nutrients. Sometimes the mat turns out to be dinoflagellates with an odd strand or two of Oscillatoria (!).

 

[taricha]

Here I am beavering away looking for nutrients to grow luxurious cyanobacteria mats and someone is selling a “probiotic” that will grow strong and healthy cyanobacteria! I am going to get me a bottle -)

Pretty cool putting your knowledge on visible light to practical use.

Did you get around to looking at the cyano under the microscope? I am curious about what types of organisms live in the mat. I find it varies depending on the nutrients. Sometimes the mat turns out to be dinoflagellates with an odd strand or two of Oscillatoria (!).

When you do find the formula to grow luxurious cyano mats on demand, you can call the bottle "Cyan-Yes!"

I actually left the tubes running to see what the final end state was - if they all end in cyano. I will get them under a microscope. But I'm going to pre guess majority oscillatoria on the chaeto filaments, with spirulina patches on the glass tubes. And possibly in the vinegar, some other organism.

 

[Dan_P]

When you do find the formula to grow luxurious cyano mats on demand, you can call the bottle "Cyan-Yes!"

I actually left the tubes running to see what the final end state was - if they all end in cyano. I will get them under a microscope. But I'm going to pre guess majority oscillatoria on the chaeto filaments, with spirulina patches on the glass tubes. And possibly in the vinegar, some other organism.

The product name might also be Cyano-Dan

 

[taricha]

I actually left the tubes running to see what the final end state was - if they all end in cyano.

Below is the state of the cultures on day 23. Compare to the cultures on day 16 in post #395 I feel like at this point they've largely run their course.

Cyano continued to increase across all cultures except the control. The chaeto-only increased significantly, almost reaching the level of the vibrant. I suspect the final end state of stagnant decomposing cultures will be cyano in nearly all cases. The vinegar treatment is the outlier here and continues to have super low cyano levels.

My tentative interpretation is that the control lacked the nutrients to support significant cyano growth. In other treatments chaeto decomposing drove the cyano growth as expected, except in vinegar treatment. Vinegar spurred bacteria that was unhelpful to cyano and grew in a surface film that locked nutrients up unavailable for cyano & associates growth. The vibrant grew cyano much like the chaeto-only, but faster because it presumably kickstarted the bacterial associates helpful to cyano.


Still need to do some microscope checking to see what actually grew.

 

[Belgian Anthias]

As do most bacteria, Cyano prefere ammonia as a nitrogen source. When using nitrate-nitrogen the growth rate is slowed down. As cyano, part of a microbial matt, are normally coverd by heterotrophs using up most ammonia one may considder there normal growth rate is based mainly on the consumption of nitrate-nitrogen and ammonium-nitrogen produced within the matt. The parameters of nitrate and phosphate in the watercolumn do not matter much ( for the cyano in the film) exempt when there is not enough.
Most cyano are able to store phosphate for 4 cel divisions which makes them competitive to other PAO ( Phosphate Acumulating Orgamisms) for other essential building materials.
As the growth rate of heterotrops controls the availabilty of oxygen and ammonia in the layers, the availabilty of usable organic carbon in the water column may play a main role in the growth rate of cyano and other photo-autotrophs.