First Time Phyto Culture Questions

Jobu183

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Hello- Couple of questions as I'm starting my phyto journey.

I'm growing nanno (front two bottles) and tetra (back bottle). I'm in my second batch, and am seeing that after ~ 5 day the culture starts to get a bit pale / yellow. Even while the culture is more green on day 4-5, I still wouldn't say that it is super dense.

Set up is 1.025sg salt water and 1ml of F2, ~30% starter culture / 70% SW in a sterilized 1 liter container.

1.) If I use my hanna test kits for nit / phos, that should tell me if I need more fertilizer correct? I.E. if my nit / phos are showing low readings, however the culture is not dense green yet, it would be a sign to add more fertilizer?

2.) The recommended density of my F2 is 1ml / liter, which is what I am dosing. I've read a few threads that people have gone a bit over this recommended dose. Wondering if anyone has had success with this in creating a darker culture.

Thanks in advance for any help. Pic attached for reference

image0 (2).jpeg
 

Js.Aqua.Project

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I would expect too much color interference from the phyto for an accurate result in a Hanna Checker (or probably most NO3/PO4 tests).

They phyto itself is a really good gauge for how much you need to add, if it is turning pale it is either time for a) more f/2 or b) your culture is crashing.
 
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Jobu183

Jobu183

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I would expect too much color interference from the phyto for an accurate result in a Hanna Checker (or probably most NO3/PO4 tests).

They phyto itself is a really good gauge for how much you need to add, if it is turning pale it is either time for a) more f/2 or b) your culture is crashing.
Appreciate your answer.

In my research, I believe crashes can be caused by either bacteria, contaminate, etc. OR by the culture being too dense, in which case I would want to split the culture rather than add more fertilizer. Is there a way to tell the difference between the two?

My issue in being new to this is that, I'm a bit unsure if my nanno is just supposed to be that lighter green color, and I should be splitting it, or if I should be adding more fertilizer.
 

Mr. Mojo Rising

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it doesn't look good to me, I would start over.

Really hard to know what went wrong... maybe you didn't start with enough phyto, maybe not enough f2, maybe something was contaminated, maybe the light too strong.... like you said, its your first time, its a learning process.
 

Js.Aqua.Project

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Appreciate your answer.

In my research, I believe crashes can be caused by either bacteria, contaminate, etc. OR by the culture being too dense, in which case I would want to split the culture rather than add more fertilizer. Is there a way to tell the difference between the two?

My issue in being new to this is that, I'm a bit unsure if my nanno is just supposed to be that lighter green color, and I should be splitting it, or if I should be adding more fertilizer.
So like @Mr. Mojo Rising said, it's a learning experience.

I will say it's harder to gauge in smaller bottles than larger ones, but on density, you're looking for it to be dense enough not to be able to see through the vessel and about the shade of a Key Lime to a Persian Lime. If it's turning brown is usually a sign of too many nutrients (but could also be a sign of contamination).

A crashing culture will usually go clear rapidly - for a large 5-10 gallon culture this can be over the course of a day or so, for smaller cultures a few hours.

One of the things I monitor is the bubbles at the top, if you're noticing a foam head forming you're most likely getting die off within the culture which will lead to bacterial contamination. I have found (for me at least) this happens when letting the culture go to long without harvesting and it getting too dense.
 
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Jobu183

Jobu183

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it doesn't look good to me, I would start over.

Really hard to know what went wrong... maybe you didn't start with enough phyto, maybe not enough f2, maybe something was contaminated, maybe the light too strong.... like you said, its your first time, its a learning process.

Is too much light a thing? I definitely have a lot of light on them, but after seeing most people using the Poseidon Reef kits, those look to have a ton of light on them, so I thought the more the better?

I will say it's harder to gauge in smaller bottles than larger ones, but on density, you're looking for it to be dense enough not to be able to see through the vessel and about the shade of a Key Lime to a Persian Lime. If it's turning brown is usually a sign of too many nutrients (but could also be a sign of contamination).

A crashing culture will usually go clear rapidly - for a large 5-10 gallon culture this can be over the course of a day or so, for smaller cultures a few hours.

One of the things I monitor is the bubbles at the top, if you're noticing a foam head forming you're most likely getting die off within the culture which will lead to bacterial contamination. I have found (for me at least) this happens when letting the culture go to long without harvesting and it getting too dense.

Yeah, mine aren't crashing that fast, I first noticed today that they looked a bit lighter, which caused me to ask for help. "Key Lime" I would say is the color that I've had in my culture, but I guess I was waiting for a more "Forest Green" type of dark green. So far the bubbles at the top have been clear and pop immediately (ie. no foam).

Trying to save at least one of these cultures as I only have about 4ml or so of my starter left and would really prefer not to have to ship another one. Added a bit more fertilizer this morning and hoping that helps. Also reduced the light to @Mr. Mojo Rising point.

Thanks both
 

Js.Aqua.Project

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Is too much light a thing? I definitely have a lot of light on them, but after seeing most people using the Poseidon Reef kits, those look to have a ton of light on them, so I thought the more the better?



Yeah, mine aren't crashing that fast, I first noticed today that they looked a bit lighter, which caused me to ask for help. "Key Lime" I would say is the color that I've had in my culture, but I guess I was waiting for a more "Forest Green" type of dark green. So far the bubbles at the top have been clear and pop immediately (ie. no foam).

Trying to save at least one of these cultures as I only have about 4ml or so of my starter left and would really prefer not to have to ship another one. Added a bit more fertilizer this morning and hoping that helps. Also reduced the light to @Mr. Mojo Rising point.

Thanks both
I have never had an issue with the light being too strong but I have had the most success running an 8 on 4off light cycle. However, some facilities run their lights 24/7 and some do a 12 on 12 off or and 18/6.
 

Mr. Mojo Rising

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Is too much light a thing? I definitely have a lot of light on them, but after seeing most people using the Poseidon Reef kits, those look to have a ton of light on them, so I thought the more the better?



Yeah, mine aren't crashing that fast, I first noticed today that they looked a bit lighter, which caused me to ask for help. "Key Lime" I would say is the color that I've had in my culture, but I guess I was waiting for a more "Forest Green" type of dark green. So far the bubbles at the top have been clear and pop immediately (ie. no foam).

Trying to save at least one of these cultures as I only have about 4ml or so of my starter left and would really prefer not to have to ship another one. Added a bit more fertilizer this morning and hoping that helps. Also reduced the light to @Mr. Mojo Rising point.

Thanks both
I've turned a few cultures yellow with bright lights, when I turned my lights way down I had better results. My research usually finds that any cheap light will work, so I just correlated the bright light to my crashed culture. But like I said, its always hard to know exactly why it crashes, but thats that I think.
 
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Jobu183

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Yeah, i'm currently running a 16/8 schedule. Going to try and rescue this batch with lower lighting, and probably trying a bit more fertilizer.

Thanks again
 

Waldek M.

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Let's say I kept the culture too long (13 days) and ended up with a very green phyto. Now when I store it in the fridge it forms a slight green precipitate. Is it suitable for administration and further breeding? Previously, the same phytoplankton did not leave any sediment, but was cultured for 7-9 days. Of course, the best solution would be to use a microscope, but I'm having trouble with that at the moment
 

DaJMasta

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I think it's likely gone (a different strain, maybe a different organism), but if you want to try and get back the original, the best solution may actually to be to dilute and reset it.

Think of it this way: your culture is guaranteed to have unintended organisms in it unless you're diligently following sterile laboratory procedures, so the game is making sure your conditions are ideal for the organism you want and less good for the others. There is some overlap so you can't fix everything, but high light and aeration tends to favor fast growing photosynthetics, so provided you're not burning them with light or ejecting then from the bottle, that's a good direction for most algae cultures. Your fertilizer quantity is on the higher side, but is pretty much in line with what I've settled on (5mL per gallon, 2.5mL of each of the two part fertilizer I use), and I think generally phyton can and will use it as quick as anything else.

The resetting can help when you run into an organism that outcompetes the phyto when grown in, but which takes longer to get established. By resetting the culture, you give the chance for the phyto to bloom quickly and use up the available nutrients, and by staying in this boom phase of the cycle by restarting the culture (take a small portion and put it in a new bottle with new fertilizer and water), you can keep the balance of organisms to favor the quick growing phyto.

In my experience, color changes are either a new strain taking hold or a crash in progress, and the way to tell which is the smell - normal smell is a different phyto strain, bad smell is a crash. If it's crashing, it's unlikely you can save it without a lot of knowledge and tools (not worth the effort too, probably), but if it's a different strain you can choose whether to keep the new one, try resetting the culture to get a bloom of the one you want, or just ditching it.

Nanno should be slightly yellower looking than tetraselmis, but not yellow looking. While in theory, adjusting light amount can be used to target the strain you want, they are all adaptable and we don't know enough about the specific intensities they do best at to be able to target one specifically. Use lighting amount changes to change overall culture growth rate, it's unlikely to be a useful tool in trying to recover a culture.
 

Waldek M.

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I think it's likely gone (a different strain, maybe a different organism), but if you want to try and get back the original, the best solution may actually to be to dilute and reset it.

Think of it this way: your culture is guaranteed to have unintended organisms in it unless you're diligently following sterile laboratory procedures, so the game is making sure your conditions are ideal for the organism you want and less good for the others. There is some overlap so you can't fix everything, but high light and aeration tends to favor fast growing photosynthetics, so provided you're not burning them with light or ejecting then from the bottle, that's a good direction for most algae cultures. Your fertilizer quantity is on the higher side, but is pretty much in line with what I've settled on (5mL per gallon, 2.5mL of each of the two part fertilizer I use), and I think generally phyton can and will use it as quick as anything else.

The resetting can help when you run into an organism that outcompetes the phyto when grown in, but which takes longer to get established. By resetting the culture, you give the chance for the phyto to bloom quickly and use up the available nutrients, and by staying in this boom phase of the cycle by restarting the culture (take a small portion and put it in a new bottle with new fertilizer and water), you can keep the balance of organisms to favor the quick growing phyto.

In my experience, color changes are either a new strain taking hold or a crash in progress, and the way to tell which is the smell - normal smell is a different phyto strain, bad smell is a crash. If it's crashing, it's unlikely you can save it without a lot of knowledge and tools (not worth the effort too, probably), but if it's a different strain you can choose whether to keep the new one, try resetting the culture to get a bloom of the one you want, or just ditching it.

Nanno should be slightly yellower looking than tetraselmis, but not yellow looking. While in theory, adjusting light amount can be used to target the strain you want, they are all adaptable and we don't know enough about the specific intensities they do best at to be able to target one specifically. Use lighting amount changes to change overall culture growth rate, it's unlikely to be a useful tool in trying to recover a culture.
The only change was the length of breeding. Lighting, aeration and nutrient (f2) all the same. Sterility is always maximum and I don't worry about it. There is no smell and it behaves identically in the bottle except for a darker color and a slight sediment. I checked 6 days after starting the breeding and everything looked normal, but unfortunately due to the multitude of duties I forgot to finish in time. I started a new breeding and I'll see how it turns out
 

DaJMasta

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You say maximum, but I think you underestimate the extremeness of plankton culturing sterilization procedures (putting your fertilizer in a starter culture flask, covering it with tin foil, and autoclaving the entire thing before innoculating it because when you pull it out of the autoclave if it's not covered, things can just settle in from the air) and how even that measure of care will never prevent all contaminants, but that is beside the point.

The organisms making your culture change color have probably been there for a while, and it's just now that they're being noticed because they've multiplied sufficiently to overtake or inhibit the nanno. That it doesn't smell means it's probably being taken over by another species rather than just killed off/infected, which is well and good, but if it's not the species you want or it has some other problem (leaves residue that's annoying to clean, settles to the bottom, foams up on the top, etc.) it may not be worth keeping regardless.

FWIW, I'd also recommend not testing for nitrate or phosphate. If you want to know the levels, make up a new bottle with no algae and measure that, but testing actually increases your risk for contamination and cross contamination 10x or more - you really don't want to be doing anything in the culture after it is setup, because anything touching the culture itself will contaminate it with something.
 

Waldek M.

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Believe me, I appreciate the importance of sterility and I know that I have preserved it, and the phytoplankton has not changed color, it is just more intense due to too long cultivation.
 

DaJMasta

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Sorry, I didn't catch it before, but my post was aimed at the OP's, I guess it was a bit of a necro.
 

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