What am i doing wrong with phyto culture?

[mrWinter]

Can anyone tell me what I'm doing wrong with my phyto culturing methods? I have read everything I can find, watched YouTube videos, and MACNA lectures, and still can't get things really going with my nanochloropsis culturing.

I have an air pump with a filter on the tubing to keep contaminants out, and I use f2 from mercer of montana. I have tried:
1) plastic containers wiped with vinegar then dried out
2) plastic containers rinsed and wiped with alcohol then dried out
3) boiling all tubing before setting up
4) glass containers that I boil the water directly in, then let cool down and add f2
5) using fresh 1.020 rodi water in sanitized containers
6) using 1.023-1.024 tank water I boil directly in container
7) very strong lighting (quantify? I dunno, but hurts to look at)
8) half the above lighting
9) another much more chill light strip
10) using magikocean as culture
11) using nano cultures from Mercer of Montana

No matter what I do nearly all the algae seems to settle out and die in short irder. The water doesn't go totally clear quickly but maybe overnight I'll see some settling on the bottom and then more and more and the culture never darkens. I did have some moderate success but it didn't last and ended up the same as above. I'm having a hard time imagining what I could be doing wrong that is introducing contaminants especially with the boil water directly in culture vessel method, also how they could be crashing things so fast. Only other thought I have of late is that I contaminated my f2 container? I wipe the outside of the container and the syringe with alcohol now before adding but maybe the contents themselves are contaminated? Other than that I don't know what else could be going wrong. It seems like nano is so easy to culture and I'm now going above requirements but no luck. I'd love any suggestions folks have. See pics below of color and settling of my two current culture attempts next to a macro grow out I have between them. Thanks any and all.

 

[ocncheffy]

You boil the saltwater? It sounds like your overcomplicating it. These are my steps:

1. Clean container with citric acid
2. Rinse with tap water
3. add 300ml of starter culter and fill with 3.5l of 1.026 saltwater freshly mixed
4. add 2ml of F2 fert.

By day 7-9 I have a nice dark culture. My lighting runs for 16 hours.

 

[Subsea]

I just had a batch of Tetraselmis from Mercer of Montana via Amazon do what happened to you.

I have successfully grown phyto without the strict protocoal you adhered to.

With respect to sterilizing/boiling water, consider using distilled water which has been boiled to get distilled.

 

[Derrick0580]

You can’t have too much agitation, maybe try cranking up your pump or getting a larger one.

 

[Js.Aqua.Project]

I agree with the previous posts, I've cultured nanno for years and unlike other phytos it doesn't require nearly as rigid a routine as you're going through.

Personally I don't use tank water in my cultures, I always used fresh mixed saltwater.

Here is my routine:
-bleach container, hand dry, then leave to air dry for 24+ hours (if you don't have 24 hours then use Prime to neutralize the bleach)
-mix fresh batch of saltwater to 1.020, add 1 drop bleach per gallon and leave to mix for 1 hour, then add Prime at 1ml/gal and leave to mix for 1 hour
-Add phyto and sanitized saltwater to sanitized container at 1:1 ratio
-Add f/2 media

Double the water volume every time it becomes too thick to see through and add appropriate amount of f/2 media

Aeration should be high enough to keep phyto in suspension but not enough to cause it to splatter up the sides of the container.

 

[mrWinter]

You boil the saltwater? It sounds like your overcomplicating it. These are my steps:

1. Clean container with citric acid
2. Rinse with tap water
3. add 300ml of starter culter and fill with 3.5l of 1.026 saltwater freshly mixed
4. add 2ml of F2 fert.

By day 7-9 I have a nice dark culture. My lighting runs for 16 hours.

Yes I was boiling tank water, I thought this would be a rather tidy way to do it, just take tank water out, sterilize it by boiling in the culture container, then add phyto and fert, then replace same tank water later back into tank now full of nanno. Same salt that came out goes back in and no mixing new water and no separate sterilization of the culture container. If I keep adding 1.020 cultures, after time I'll be raising salinity in my tank.

I like that you were adding 300ml of culture to 3.5 l of fresh water and having success, this sounds much better than the repeated doubling. I never understood why adding a little to a lot with appropriate amount of fertilizer didn't work, but I'm going to try going back to the 1:1 for now until I have success with that.

I just had a batch of Tetraselmis from Mercer of Montana via Amazon do what happened to you.

I have successfully grown phyto without the strict protocoal you adhered to.

With respect to sterilizing/boiling water, consider using distilled water which has been boiled to get distilled.

Thanks for the reply, I"m wondering if this is part of my issue, maybe it spent too much time on a shelf in an amazon warehouse or got too hot in transit, not sure.

 

[mrWinter]

You can’t have too much agitation, maybe try cranking up your pump or getting a larger one.

I could definitely bubble more vigorously, I had it going at a couple bubbles a second. I'll try that, thanks.

 

[Pistondog]

Do not use tank water. Try a heavier start dose, like 25% seed plankton.
Fwiw, more to start seems higher chance of success.

 

[Mr. Mojo Rising]

Personally, I see 2 things I would do different:

Does not need very strong lighting at all, I would say that very weak light is better.

I would try to find a single phyto strain, like nanno for example, instead of trying to grow corporate phyto from a bottle like magicocean.

I think you are going way overboard with all that sanitizing, but if you use the wrong light and wrong starter culture, then it doesn't matter.

And don't use tank water like mentioned above, use newly mixed saltwater. All that sanitizing is pointless if you are contaminating the culture with algae from your tank.

 

[Derrick0580]

I could definitely bubble more vigorously, I had it going at a couple bubbles a second. I'll try that, thanks.

Couple bubbles a second is for copepods