Dino ID + Treatment

[JonasRoman]

If you have time for reading, I'd like to hear what you and others think about this and what they describe about N and P's role:

(Nature) https://doi.org/10.1038/s41598-023-41339-3

Excellent, will read later

 

[martin.a]

If you have time for reading, I'd like to hear what you and others think about this and what they describe about N and P's role:

(Nature) https://doi.org/10.1038/s41598-023-41339-3

Quick and dirty by no expert:

Well, they basically show what the title of that paper says: P deficiency triggers sexual reproduction in dinos. Interestingly, it happens only when there is no N deficiency. They did not observe the same response when both N and P were bottomed out. That's actually interesting. At least to me.

Additionally, they claim: "Nitrate uptake was significantly impaired in the P-deficient culture [...] Phosphate uptake was significantly reduced in the N-deprived culture [...]". What a surprise ;-)

BTW, I can't believe Sci. Reports (not Nature!) allowed them to use the word "significantly" in a context other than statistical significance. Funny that they did not do even simple stats, as Figs. 1-3 look like they were done in R with ggplot2.

The take-home message is: when there is plenty of N and P, the dino population grows rapidly through vegetative reproduction. When P is deficient but N remains available, they switch to sexual reproduction and population growth stalls.

Anyway, the main limitation is that the analysis was done on Prorocentrum cordatum only. So, the conclusions are valid only for this species and cannot be generalized to other dinoflagellates.

 

[martin.a]

My comments are to the point and might sound harsh if you're looking at other solutions or conflict with your views . If you search on R2R regarding dinos, I'm not saying anything new and I'm only sharing my perspective and comments.

Apologies if I come off offensive or confrontational. Just offering a solution to resolve your issues ASAP given I've experienced it many times and resolved it the same way.

I do appreciate you sharing your expirience. If anybody should apologise here for straight to point language it would be me. Seems I have a lot to say, but not much practical experience.

I dose P and N to keep them above zero. But this is counter-intuitive to me: Dinos need both to proliferate. So basically I am feeding the beast. How dinos outbreak can be triggered by lack of nutrients? What would they use?

 

[JWsticks]

My comments are to the point and might sound harsh if you're looking at other solutions or conflict with your views . If you search on R2R regarding dinos, I'm not saying anything new and I'm only sharing my perspective and comments.

Apologies if I come off offensive or confrontational. Just offering a solution to resolve your issues ASAP given I've experienced it many times and resolved it the same way.

I do appreciate you sharing your expirience. If anybody should apologise here for straight to point language it would be me. Seems I have a lot to say, but not much practical experience.

I dose P and N to keep them above zero. But this is counter-intuitive to me: Dinos need both to proliferate. So basically I am feeding the beast. How dinos outbreak can be triggered by lack of nutrients? What would they use?

You have a nice build and it is frustrating to see it with dinos. Maintain NO3 at 2.5 and phosphate around the 0.10 - 0.20. This step is critical and keeping a log will help you through the next occurrence regarding how much PO4 to dose and time it takes to overcome it. With every other day testing and dosing accordingly, dinos should be gone after 1-2 weeks. I would even go stir up the sand bed ensuring everything is exposed to PO4.

I'm almost done with dinos in my tank and can see the snots releasing itself into the orderflow and the white rocks now have dark green algae growing on them instead of snots.

Limiting the photoperiod would also help with growth given dinos are photosynthetic.

 

[JWsticks]

The tank has been up for 4 months (120g 4x2x2), but I started with ocean live rock and dry rock that I seeded for about a year in a brute.

Fish:
Flame Hawk
Yellow Tang
Tomini Tang
One Spot Fox Face
PJ Cardinal
Banded Sleeper Goby (for now... he is way too messy)

Feed 1/2 sheet of nori daily + assortment of pellets.

As for corals, I just have a few hammers currently. They are doing great. I will hold off on adding more until this issue has been resolved.

Ah, you're still in the early microbiome stage. Hammers won't be effected too much unless the dinos are toxic and the outbreak is bad. I definitely recommend the hydrogen peroxide treatment. Nothing in the tank would take it hard and it's a pretty gentile treatment. It will take about a month to clear up.

I've seen dry rock both absorb and leech a lot of nutrients. It definitely take a while to stabilize. I'd recommend stopping all other dosing other than your big 3 and stop water changes. From my experience, dinos take over from instability more than anything else.

Also as soon as the dinos die off, you will probably get hair algae. The tangs and foxface should help keep it at bay. The best combat to the algaes is a good layer of coraline algae.

Thanks for the tips! Luckily the rocks are unaffected and currently covered in coraline. This is only on my sand.

Rocks already at equilibrium with PO4 hence why rocks are not affected. Take out the sand and treat it with PO4 if you want to be aggressive or continue to dose nutrients.

After dinos, I would suggest a refugium with a strong LED to manage nutrients in a balanced manner. Vodka is better at reducing NO3 vs PO4 and you'll prolly need lanthanum or gfo to lower PO4 if you're feeding heavy.

 

[EnterName]

I do not believe the po4 absorbed in sand causes dino. That is fixed in some part, insolubable. One part is in equilibrium but always go back and forth and is not exclusively addressed to dino.

This is just a theory and nothing I ever bothered to verify, but: we are talking about benthic dinoflagellates, which sit on the substrate or on rocks and form mats. It wouldn't surprise me if these mats had an impact on the water chemistry below them, making the bound phosphate available to dinos through local pH changes or something similar. Cyanobacteria are already known to make use of this phenomenon (Wood et al. 2015), so it's possible dinos have similar abilities.

How dinos outbreak can be triggered by lack of nutrients? What would they use?

I'm not too sure about the exact trigger, but we have some indicators of what they use:

Ostreopsis cf. ovata (a potential candidate for the dinos we actually find in our tanks) grows just as well on a wide range of dissolved organic phosphorus compounds as on inorganic phosphate (Ellwood et al. 2020). Its biofilms show phosphatase activity, meaning enzymes that cut phosphate off organic molecules. In other words: They have access to phosphate we can't measure at home and may also have high-affinity uptake mechanisms that allow them to drain phosphate from the water at lower levels than many other organisms.

It's not completely abnormal for natural coral reefs to have something like 0.005 ppm PO₄ without everything starving to death, so the uptake limit of these organisms is definitely lower than our testing methods allow us to measure. They might not reach optimal reproduction rates at this point anymore, but it won't cause mass starvation on its own either.

There are also heterotrophic feeding capabilities in dinoflagellates, but for the few genera that frequently cause trouble in reef tanks, the reports seem quite limited:

Prorocentrum (planktonic species)	Not bloom-forming in our tanks. Solid proof of phagotrophy.
Prorocentrum (benthic species)	"Stained [food] vacuoles were not registered in the 330 Prorocentrum spp. cells observed."
Coolia	"only [...] 2.2% of 134 [...] cells presented evidence of phagotrophy [...]"
Ostreopsis	"only 0.4% of 1195 [...] cells presented evidence of phagotrophy [...]", but Lee & Park, 2018 found up to 29.5%.
Amphidinium (colorless species)	Not bloom-forming in our tanks. Needs food, phagotrophy essential.
Amphidinium (pigmented species)	Never really measured. A. carterae has been seen ingesting a small diatom species, but the ingestion rate wasn't measured.

I have just ordered suitable stains to check for myself whether I can detect phagotrophic feeding in my experiments by detecting food vacuoles in the cells I have available... can't hurt to double check if this stuff applies to reef tanks.

References:

• Almada et al. 2017 - "Investigation of phagotrophy in natural assemblages of the benthic dinoflagellates Ostreopsis, Prorocentrum and Coolia."
• Mitra et al. 2023 - "The Mixoplankton Database (MDB)"
• Ellwood et al. 2020 - "Growth and phosphatase activities of Ostreopsis cf. ovata biofilms supplied with diverse dissolved organic phosphorus (DOP) compounds"
• Lee & Park, 2018 - "Genetic Analyses of the rbcL and psaA Genes From Single Cells Demonstrate a Rhodophyte Origin of the Prey in the Toxic Benthic Dinoflagellate Ostreopsis"
• Wood et al. 2015 - "Entrapped Sediments as a Source of Phosphorus in Epilithic Cyanobacterial Proliferations in Low Nutrient Rivers"

 

[Mas0n]

Alrighty!

Noticing a few things today, sand and glass are still super ugly. BUT, it seems like my sleeper goby has been more interested in the sand and filtering it. He seemed to slow down with that once the dinos appeared in full force. My yellow tang is also picking a lot more. I can see some "hair algae" like stuff forming on the sand and walls, which is great.

You can see all the little places where he takes a bite towards the front, and towards the midground of the photo the sand is basically half dino half white.


As for Nutrients:
30.2 No3
.05 Po4

Dosed an additional 20ml Neophos.

Two things stand out to me.

First, after dosing 20ml Neophos I expected a greater increase. So its either expired and not effective or my consumption is just that high.

Secondly, my no3 tested at 18.4 yesterday... this is likely a testing outlier and my nitrates were closer to the mid 20s. I am noticing some inconsistency across my nitrate testing... But the curves kind of match my po4.

 

[mcarroll]

I do appreciate you sharing your expirience. If anybody should apologise here for straight to point language it would be me. Seems I have a lot to say, but not much practical experience.

I dose P and N to keep them above zero. But this is counter-intuitive to me: Dinos need both to proliferate. So basically I am feeding the beast. How dinos outbreak can be triggered by lack of nutrients? What would they use?

There is still a lazy through-line in this conversation....

We're conflating "real" dino blooms, aka the tank-wrecking toxic kind, with "algae dino's" that just seem to grow like algae in a relatively harmless manner.

There's no reason both should require the same treatment.

Another aspect of that is in many ways the P is not for the dino's – we're also hoping to encourage less hardy but equally important (and maybe moer beneficial) periphyton members to populate the tank.....and that's unlikely under starvation conditions.

So keeping nutrients replete is part of an overall strategy for a healthy growing reef, not just a specific thing to squelch dino's.

How dinos outbreak can be triggered by lack of nutrients? What would they use?

They are triggered (by starvation signals) to eat bacteria instead of photosynthesizing.

Bacteria have less problems accumulating P even under limited conditions so they are a great snack, and are practically unlimited in the tank.....so dino's bloom on this huge food source.

https://reefsuccess.com/2016/10/17/...survival-strategy-during-nutrient-limitation/

I don't think most folks account for this possibility....I know when I ran across this article the first time I had to do a triple-take......"algae eat whaaaaa...?"..."algae 'eat'...?" Okay.

First, after dosing 20ml Neophos I expected a greater increase. So its either expired and not effective or my consumption is just that high.

It doesn't expire, so consumption is just that high.

Secondly, my no3 tested at 18.4 yesterday... this is likely a testing outlier and my nitrates were closer to the mid 20s. I am noticing some inconsistency across my nitrate testing... But the curves kind of match my po4.

I wonder if you take a sample and test it in triplicate how all three results look next to each other. I'd hope for pretty good parity between them.....if not I'd think maybe something in the process isn't happening right. I do this whenever I get outlier/non-predicted results just to make sure I'm doing it right and the test itself isn't weird/exppired for some reason.

 

[JWsticks]

Alrighty!

Noticing a few things today, sand and glass are still super ugly. BUT, it seems like my sleeper goby has been more interested in the sand and filtering it. He seemed to slow down with that once the dinos appeared in full force. My yellow tang is also picking a lot more. I can see some "hair algae" like stuff forming on the sand and walls, which is great.

You can see all the little places where he takes a bite towards the front, and towards the midground of the photo the sand is basically half dino half white.


As for Nutrients:
30.2 No3
.05 Po4

Dosed an additional 20ml Neophos.

Two things stand out to me.

First, after dosing 20ml Neophos I expected a greater increase. So its either expired and not effective or my consumption is just that high.

Secondly, my no3 tested at 18.4 yesterday... this is likely a testing outlier and my nitrates were closer to the mid 20s. I am noticing some inconsistency across my nitrate testing... But the curves kind of match my po4.

Dosing PO4 initially was uncomfortable and counterintuitive for me as I was always exporting it.

As for your daily dosing amount, I would suggest to dose 0.10 PO4 daily. You'll prolly have to go through a bottle of phosphate.

I've been dosing .11 PO4 daily and one day I accidentally poured a bit too much and spiked it where I only dosed half dose for that day.

Most of the brown dinos on the rocks have now turned into dark green algae however still see some long snots on the back glass. Feels like I'm almost near the finish line.

I think mixing the sandbed manually would help expose the sand to PO4 as sand can create areas where flow cannot reach. @mcarroll may have more experience on this.

 

[JonasRoman]

just a quick one, is reading the article now. Lets say its true that low P induced sexual reproduction (but still necessarily not more visible dinos, just more DNA), and now you add P, and those now more cells explode in vegetative phase and you just fuel them. Thats why I mean when I believe its contraproductive to dose P. Here is from the article (in the abstract section); "Subsequent supplementation with phosphate stimulated meiosis and led to a noticeable increase in the 4C cell number (dividing zygotes"

Will do a deeper reding later today.

Jonas

 

[JonasRoman]

I have red results and discussion. I interpet the article in this way:

1) Dino grows more unlimited in a vegetative way when both N and P exists.
"The P-defcient culture reached a plateau in 15 days with
maximum biomass 1.66× 105
cell mL–1, while control cells continued to divide until day 18"
dinofagellate blooms start and develop in favorable conditions, when
nutrients are abundant. At this period of time, dinofagellates reproduce vegetatively
2) Dino consumes quiet much N and P as long as both exists.
the control, nitrate and phosphate concentrations gradually decreased from 630 to 72 µM and from 37 to 2µM.
3) If P is totally depleted, N consumption stops, and also dino goes into a sexual reproduction phase (and cystic) as a strategy to survive when P is deficient. Dino now do not vegetatively growth.
nutrient defciency, especially N and
P limitations, are considered to be the most efective factors to induce sexual reproduction
5) If we dose P now, the dino leaves the sexual reproduction stage and go back to vegetative and start to thrive again.
addition of nutrients can stimulate meiosis and return to the initial vegetative state.

Jonas

 

[mcarroll]

It seems worth clarifying that during a tank-wrecking-toxic-dinno-bloom (which is not happening in this thread) the idea is to get the tank back to balance....eradicating dino's isn't really the mission. (Returning them to peaceful non-toxic epiphytic behavior is the mission.)

So whether cell counts ever go up during nutient enhancement, I'm not sure....but you can definitely see them drop form initial levels where they dominate microscope slides to the point of monopoly down to much lower levels, often undetectable. (Nobody scans the water column, so no data there.....miraculous I was able to get so many folks to buy and use microscopes!!)

The idea is for them to return to an epiphytic lifestyle – clinging to algae, etc.

Here, we seem to be dealing with other types of dino blooms that aren't toxic (eg. non-toxic, not as toxic, whatever), and seem to be more of a visual nuisance than they are a real problem.

The goal in this case isn't as clear, but seems like it might purely be about reducing cell counts so the dino's are no longer visually offensive.

If so, then whether they are veg or sex is much less relevant....we're trying to "make them happy epiphytes again" not make them disappear, which is a much tougher row to hoe. Better to encourage other things to grow with the dino's and just be happy the dino's aren't blooming anymore.

Shifting subjects slightly..... Did anyone else have Nature recommend the following article after the one I linked? I just got it recommended when I was reviewing the veg/sex link again. This article is also pretty interesting. They test dino's (not our genus) as well as diatoms through a regime of nutrient depletion with the goal of predicting the end of a phyto bloom metabolically...and project that this may be useful across phytoplankton types. (They're working on a patent for testing this.) We aren't going to be testing like this, but some details of the experiment still seem like they might be useful. I also think this one probably speaks more directly to the trend @JonasRoman is trying to detail.
"Chronological age-related metaboiome responses in the dinoflagellate Karenia mikimotoi, can predict future bloom demise"
(nature) https://doi.org/10.1038/s42003-023-04646-z

 

[Mas0n]

It seems worth clarifying that during a tank-wrecking-toxic-dinno-bloom (which is not happening in this thread) the idea is to get the tank back to balance....eradicating dino's isn't really the mission. (Returning them to peaceful non-toxic epiphytic behavior is the mission.)

So whether cell counts ever go up during nutient enhancement, I'm not sure....but you can definitely see them drop form initial levels where they dominate microscope slides to the point of monopoly down to much lower levels, often undetectable. (Nobody scans the water column, so no data there.....miraculous I was able to get so many folks to buy and use microscopes!!)

The idea is for them to return to an epiphytic lifestyle – clinging to algae, etc.

Here, we seem to be dealing with other types of dino blooms that aren't toxic (eg. non-toxic, not as toxic, whatever), and seem to be more of a visual nuisance than they are a real problem.

The goal in this case isn't as clear, but seems like it might purely be about reducing cell counts so the dino's are no longer visually offensive.

If so, then whether they are veg or sex is much less relevant....we're trying to "make them happy epiphytes again" not make them disappear, which is a much tougher row to hoe. Better to encourage other things to grow with the dino's and just be happy the dino's aren't blooming anymore.

Shifting subjects slightly..... Did anyone else have Nature recommend the following article after the one I linked? I just got it recommended when I was reviewing the veg/sex link again. This article is also pretty interesting. They test dino's (not our genus) as well as diatoms through a regime of nutrient depletion with the goal of predicting the end of a phyto bloom metabolically...and project that this may be useful across phytoplankton types. (They're working on a patent for testing this.) We aren't going to be testing like this, but some details of the experiment still seem like they might be useful. I also think this one probably speaks more directly to the trend @JonasRoman is trying to detail.
"Chronological age-related metaboiome responses in the dinoflagellate Karenia mikimotoi, can predict future bloom demise"
(nature) https://doi.org/10.1038/s42003-023-04646-z

Nailed it. They are a visual nuisance in my case. I am trying to reel their population in by encouraging easier to deal with nuisance algae to grow, which likely won't happen if my po4 and no3 bottom out.

I'm cool with Cyano, GHA, and Diatoms!

 

[RobertK]

Nobody scans the water column

Not true, see my post #32. @EnterName taught me the technique, it works.

 

[mcarroll]

Not true, see my post #32. @EnterName taught me the technique, it works.

I appreciate that....2 of us among the [thousands?|millions?] of reefers I suspect.

I didn't say I'm GLAD nobody tests.....I wish everyone did. I don't even know how.

Have you written up a thread to promote the idea?

Doing a thread worked for me and promoting microscope use as you see here and by the "rampant microscope use" in the dino thread that resulted:
https://www.reef2reef.com/threads/selecting-a-microscope.308826/
https://www.reef2reef.com/threads/dinoflagellates-–-are-you-tired-of-battling-altogether.293318/

Early on in the thread, I made ID via microscope pretty much a precondition for participating in finding the solution(s). (All we had before was the generic "shake test" for dino's. Good, but not nearly as good as pics and movies.)

 

[RobertK]

I appreciate that....2 of us among the [thousands?|millions?] of reefers I suspect.

I didn't say I'm GLAD nobody tests.....I wish everyone did. I don't even know how.

Have you written up a thread to promote the idea?

Doing a thread worked for me and promoting microscope use as you see here and by the "rampant microscope use" in the dino thread that resulted:
https://www.reef2reef.com/threads/selecting-a-microscope.308826/
https://www.reef2reef.com/threads/dinoflagellates-–-are-you-tired-of-battling-altogether.293318/

Early on in the thread, I made ID via microscope pretty much a precondition for participating in finding the solution(s). (All we had before was the generic "shake test" for dino's. Good, but not nearly as good as pics and movies.)

Nope, haven't written up a thread, but might. When I first got the sieve I just tried running it back and forth in the water and didn't collect much. @EnterName suggested I use a pitcher to pour water through the sieve, which made a big difference. After pouring a bunch of water through the sieve, a few drops are held in the bottom of the sieve by surface tension - thats what I collect with a pipette and transfer to a slide for microscopy. Works great!

 

[mcarroll]

Nope, haven't written up a thread, but might. When I first got the sieve I just tried running it back and forth in the water and didn't collect much. @EnterName suggested I use a pitcher to pour water through the sieve, which made a big difference. After pouring a bunch of water through the sieve, a few drops are held in the bottom of the sieve by surface tension - thats what I collect with a pipette and transfer to a slide for microscopy. Works great!

Excellent! Sounds to me like something that shouldn't be a secret anymore. ;)

If you (or @EnterName, etc) make a thread, I'll be there to help promote it (with more posts) for my part.