nannochloropsis phytoplankton culture (questions)

[phyto4life]

So far I started culturing nannochloropsis phytoplankton indoors using the petri dish method inwhich will be used to add directly to my saltwater tank that is at 1.024

I added a little less then 1/4 cup of rodi water that was boiled in the microwave for 1 min then I let it cool to room temp and added it to the petri dish then placed it under 2 normal output plant light's for 24 hours

where do I go from here?

I read that I should use the q-tip and add the entire dish to 1 liter of water under a gentle rolling boil and leave for 12-24 hours

my question is do I add this to fresh water or saltwater?

because ultimately I want the culture to be saltwater @ 1.020 ppm

and should I give the algae a break for a time period between adding the dish that was being lit for 24 hours to be lit in the 1L with flow for 16 hours?

I'll have more question's as I go

[hillbillyreefer]

I started out doing it that way too. Lasted for about a month. Now when I restart cultures I just swab the top of the petri dish, and swish the Qtip around in the nsw, in a clean container with fert. Turn on the lights and bubble the crap out of it. I use saltwater of around 1.017 for the batches, some of the water evaporates as the brew matures. I've never tested it when it's done, never worried about it. When I need to harvest I turn off the air for a day or two and let it settle. Pour off the top 1/2 or so, collect the rest for use, fill the container back up with nsw, fertilize and turn the air back on. Haven't had a culture crash yet, been growing for almost 2 years. I have on a couple of occasions shut down and replaced all the equipment and restarted though. The phyto is grown in my basement which is cold around 15 C. ATM I run an 18 hr lights on cycle. I have run with 24 hr on and haven't seen any detriment to the cultures. I'm know this isn't the recommended method of culturing, but it works for me and my lazy style.

Now to keep rotifers going, haven't had much luck with them, lol.

[phyto4life]

I posted early. tomorrow at 3pm I will have the petri dish with water under light for 24 hour's

from here you say to add it straight to a salinity of around 1.017 ppm with fertilizer and bubble the heck out of it in which my pump can do because it is set to bubble a 1 gallon

my temp is probably around 70-75F

my question is how much fertilizer and what amount of pre mixed saltwater/ liter/gallon etc to use right away?

the bottle from f.a.f liquid says 1/4 tsp for 1 G @ f/2 level and double that for full strength

should I go full strength fertilizer right away?

and the petri dish says 1 liter for the entire dish to inoculate should I go more or the recommended amount at first right away?

ultimately I'll be doing 1G cultures x 8

I have a second dish so i'm open to what ever suggestions

[hillbillyreefer]

Originally Posted by phyto4life

I posted early. tomorrow at 3pm I will have the petri dish with water under light for 24 hour's

from here you say to add it straight to a salinity of around 1.017 ppm with fertilizer and bubble the heck out of it in which my pump can do because it is set to bubble a 1 gallon

That is what I do.

my temp is probably around 70-75F

I don't think temp is a large factor, with higher temps you may get faster growth. I'm not sure, never worried about it, researched it or experimented.

my question is how much fertilizer and what amount of pre mixed saltwater/ liter/gallon etc to use right away?

the bottle from f.a.f liquid says 1/4 tsp for 1 G @ f/2 level and double that for full strength

should I go full strength fertilizer right away?

I would, it should help establish the best culture you possibly can.

and the petri dish says 1 liter for the entire dish to inoculate should I go more or the recommended amount at first right away?

I would dilute it, cultures are expensive. They will multiply rapidly. My qtip swab starts take about 3 weeks to become useful feed. When I start them they look like perfectly clear water.

ultimately I'll be doing 1G cultures x 8

I have a second dish so i'm open to what ever suggestions

Start as many jugs as you want with the first dish. Save the second one for an emergency. If you have multiple cultures of the same species going it is highly unlikely you will lose them all to a crash at the same time. You can always restart a jug with phyto from another culture and still have an emergency backup dish. It really isn't difficult. I was overwhelmed by the info when I started too. It really is as easy as you want to make it.

I forgot to mention, I use miracle grow for fertilizer. The one with the smallest first and second numbers I can find. I started out using 5 ml in 2l of culture now I use 3 ml and haven't seen much difference in the cultures under the microscope. I haven't noticed any problems with this fertilizer in the tanks. I have been toying with the idea of filtering the phyto out of the culture water, and putting it into NSW but haven't done this yet.

Hope that helps.

[phyto4life]

thanks

I'll start 2 x 1G going with 1/4 tsp of fertilizer and i'm thinking of splitting it after 4 day's

after that when I split them again I might leave them a little longer and add more fertilizer

do you use a density stick and if so do u know the density I would be going for that will leave vary little fertilizer with the max amount of phyto cell's?

or a shade of green?

[hillbillyreefer]

I have enough cultures going that I run them until I need them. Usually 3 or more weeks. They are deep green in that time. I don't have a density meter. If I'm really curious I'll look at them under the microscope. Someday I might pickup a density meter, but I much prefer to keep things simple. I've too many other things going on to worry about phyto too much.

[phyto4life]

everything seems to be doing good and growing

it's good to know on how dense and dark green a culture can get

[rayjay]

I don't use the disk's any more, I buy liquid starter from ReefCrew.
While I grow a lot of nanno, I no longer use it for my tank. Instead, I feed rotifers to my tank about an hour or more after lights out and see better results, especially with corals like the acans.
I no longer sterilize the water I use for phyto culture but when I did, I used to add about 8 to 10 drops of bleach to a 2L pop bottle, fill with tap water, and let sit for at least 24 hours. Then, I add ClorAm-X (or any dechlorinator) until the chlorine is neutralized and then dump half into another sterilized bottle and top both bottles up with full strength nanno that is already cultured.
It was too much hassel to keep sterilizing the water at the volumes I use now so I just take my chances on crashes and if it happens, just restart.
CULTURING PHYTO THE WAY I DO IT NOW

[phyto4life]

Thanks Rayjay good info

I'm using 1/2 tsp in a 1G of fertilizer but just started it with half a disc and will split the culture when it get's greener in 4 day's then bump up the fertalizer to 1 tsp hoping to go 6 days before splitting on a regular schedule

should I be aiming for 2 tsp in a 1G in the long run?

how long before I split?

[shrimpchips]

Do you guys refertilize after a certain point or number of days? Right now I'm only adding ferts in whenever I do a split.