Culture Journal, Species: Spirulina major

General
Species:  Spirulina major
Species description: Blue-green algae (Cyanobacteria), spiralled trichomes 
Culture source (link if possible):  Carolina Biological supply
If algae, CCMP # (Optional):  1475/3
http://ccmp.bigelow.edu/
Culture Establishment Date:  4/17/2013
Continuation Date:  

Culturing Vessel Details
Salinity: 1.020
Temperature: Room temperature
pH:  Not measured

Vessel description:  500mL plastic container
Lighting description:  Compact fluorescent bulb
Lighting cycle:  Constant
Aeration description:  Slow bubble

Methodologies
Split methodology: During early culture, the thin film-like growth is sucked off the bottom with a bulb pipet.

Culture medium description: 
Sterilized, fertilized water.  Fertilization is 1/2mL per L of water using FAF MicroAlgae Grow.  Sterilization is done using bleach and sodium thiosulfate. Fertilization is 1/2mL per L of water using FAF MicroAlgae Grow into sterilized ASW.  Sterilization is done using 1/2mL per liter of bleach for at least an hour and 1.5mL per liter of 1M sodium thiosulfate. 
Cell count:
 (if known)

Reference links:  
http://www.algaebase.org/search/species/detail/?species_id=24748
Additional Information
(No Pictures or Videos in the Section Please)
Notes: 
Water is sterilized using bleach, and dechlorinated using sodium thiosulfate.
 
This culture seemed to do best with near constant light (but not very bright and directly on it) and very slow bubbling.  It also did pretty well growing in petri dishes with no flow at all.  Eventually I shoved the cultures to the side one day when I needed the spot they were in for a bit and just forgot to move them back and they died out over a week or so in the dark 


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I am starting this culture with hopes that it will be a settlement substrate for Bursatella leachii veligers.
 
The culture came from www.carolina.com, was inexpensive and shipped quickly.  The downside is you really don't get a lot to start with, essentially just a test tube.  I knew this going in, so I ordered two right away, to be on the "safe" side.

 
Here each is in the 500mL containers:


 
I wanted to see what the golden yellow stuff was since it looked not at all like a "blue green algae," so I took a bit, smooshed it on a slide a bit so it wasn't just a clump and threw it under the microscope:


At lower magnifications the gold stuff just looked like debris, and I thought maybe it was some sort of substrate that the spirulina attaches to in culture (I really have no idea what I am doing here, so this will be interesting).

 
Higher magnifications show that the gold stuff also spirals:


So now I think it may be dying/old cells.  Still not certain, of course, since I don't really know what I'm doing yet...
 
Here are some more pictures because it was really fun to look at this under the microscope


In these last two pictures you can see smaller cell strings.  If my laptop wouldn't shut down when I tried to take a video, I would have for this because the small, single cell fragments were cruising around a bit. I don't know if this was actually them moving on their own, or just a by-product of evaporation off the slide, but it was pretty interesting.


 
Currently, these containers are on a shelf above the light at my culture station.  This is not their permanent home as it will be too high light for it to thrive (southern facing window and what not).  Since it is rainy/thunderstorming all day today, I knew I could squeak by with just putting them there last night, but tonight I will need to move them.  I think they will go by the veliger station, but behind the light so they get constant, but low, light conditions.  The constant light may be important, at least for the next few weeks, because a quick search last night showed that other Spirulina sp. cultures would lose up to 35% of biomass during the night.  Right now I need to get as much of this as I can until I determine if I can or can't use it for the purpose I need.
 
 

Very cool!  Nice to see somebody doing something out of the "ordinary".  Keep us updated, please.

I will
 
I wonder if I should break up the clumps.  I have no idea if the clumping is a by-product of the shipping, or if that is the preferred/better way for it to grow.  Perhaps one container I will break it up, and the other I will leave it clumped....  
 
I plan to do quite a bit of reading about culturing Spirulina in the next couple days.  So far most of what I have found has been for people growing it to eat it...but the culturing techniques should still be relevant I would think.

A quick update. The two cultures have been in about 150 mL of fertilizer water (FAF MicroGae grow). One container I left as a clump, the other I busted up a bit using just the air line (so it was still a bit clumpy, just smaller clumps)

The container that I broke up a bit actually seems to have a "colony" starting to grow on the bottom

You can see it near the center of the container.

So I have now busted up big clump in the other container and mixed both containers with the airline. I have a week before I will start to need this, but I doubt I will have much to work with at this rate!

The cultures have been receiving continuous light from the veliger culture station, but are positioned behind the light, so it's more ambient than direct.

One of the two culture bottles was harvested last night (to be used for veliger settlement experiments).  There was more in there than I originally thought.  I harvested by taking a bulb pipet and just sucking the thin, film layer off the bottom.  Nearly the entire bottom of the container had a thin, greenish layer, so I was pleased.  I also smashed the ball of Spirulina that has been sort of growing in there to get some more small pieces floating around and hopefully colonizing.  The second bottle was not touched.
 
More water and fertilizer will be added today, probably about 250mL of water and 100uL of F/2.

One of the bottles, the one that I screwed around with the most to use for the veligers has a nice layer of spirulina on the bottom.


Ramping this culture up has been slow, but it is happening it seems...