Culture Journal, Species: Isochrysis galbana

Cultures started. Picture from 7/24. Iso is 2 jars on right. 2nd from right started on 7/21, end jar started 7/24.




Same 2 jars on 7/28. Left appears same as it did within a few days of starting. Jar on right is taking a greenish hue and there is green algae on bottom of jar. Will be dumping this jar as it appears I contaminated with my Tet culture or a wild micro algae.

Due to concerns with cultures not getting as dark as others I have seen I am starting a new jar. This time I used plastic ex-mayo jar as I was concerned the thick canning jar glass was blocking too much light. Due to not being able to boil the plastic I used .5ml of bleach and then added .75ml of Prime (all I had on hand) to neutralize the chlorine. This jar was started from my original split which is continuing to stay brown but is not getting any darker.



Here is the remainder of the original culture. No green on the bottom so I am assuming it still pure.

Concerns and questions -

1. On my original split which isn't getting coffee colored, more of a green tea I use a T8 bulb. On two additional cultures I am using a T12 fixture. One was from original split and the other was from remaining iso in the bottle from the conference. I had been keeping it in the fridge and it was brown but stressed. Would the lighting be limiting and stressing the cultures?

2. As you can see both cultures below are green or turning green. I grabbed the original split and started the more brown one on a separate air system across the room. The second jar was started from the stressed culture and is completely green. Do stressed cultures turn green? I boiled the heck out of the water and really would not expect
them to be contaminated because the original split is not appearing to be and I didn't even handle the Tet jar when I split these.

As a note for the new plastic jar culture I also turned the fixture vertical so that the jar is receiving more light along the entire length of the jar.

Will inoculating a new culture with too little algae cause it to stress and/or not flourish?

Daniel,
I've read that having too little inoculate can cause a culture to struggle and allow contaminants to take over. It sounds like you thoroughly sterilized the culture media, but there could have been some chlorella or tet cells in the inoculate. I lost my last t-iso to some sort of green algae. The current one you gave me is fairing much better, so let me know if you need some.

Someone here also had a contaminated iso culture that they "purified" by letting it sit without aeration, with the concept that non-motile algae settles to the bottom, and the motile iso could then be decanted off the top (at least I think I read that in this forum).

~~~~~~~~~~~~~
Ken

Thanks for the info Ken. I will do a full split of the original tonight then. I had been stingy with the algae for fear of using up my clean culture. I will just do it and have no fear!

Putting Isochrysis in the fridge is really not a good idea.  It doesn't like cool temps, and will start dying. It is unlike Nannochloropsis which will tolerate cold quite well.  I keep Iso out of the fridge, and use it up as fast as I grow it.
 
If you started later cultures from refrigerated stock, that is probably why they did not do well.  Just a thought.

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Thanks Kathy. The last picture of the green culture was started from a refrigerated sample. I should have researched that - your advice will pay off as I had planned on saving a portion of each split in the fridge to feed my Parvo with. I don't have anything to raise at this point so I will have way more algae than copepods.

Not a bad place to be.

check out Kathy's Clowns, llc website:
http://kathysclowns.com
Captive bred clownfish and more
(Wholesale to the trade.)

Well, the plastic container was a fail. It stayed the same color for several days then suddenly went clear. I won't be using Prime anymore either. I got a bottle of sodium thiosulfate at the pool store so I will be switching to a chlorination sterilization over the microwave.

Below is the first true spilt of the phyto cultures. I used a full 25%-30% of the mature culture for the new start. I also added a second small light fixture to the opposite side so illumination should be much better. I am hoping for that nice chocolate color for the Iso.

Daniel, how are these cultures coming along?

~~~~~~~~~~~~~
Ken

Hi Ken.

Doing ok so far. The split above darkened up to the same light tea color and no green. So while no chocolate color at least I am keeping it going. Now I need to ramp up to multiple cultures to make it easier and safer to feed the Parvo I have too.

This iso is tricky stuff. My Tet just cruises along but the iso....well I am learning a lot. I split 50% of my culture into a 1/2gallon plastic container. The jars were a pain to pour and I didn't have lids for them. Bad news is the new split now has a greenish tint to it. Luckily I had not used the other half of original to feed as planned so I now added it to a new culture of about a quart volume. I think the larger volume diluted my start too much on the 1/2g. The other factor is with the increased outdoor temps the room they are in has increased about 5deg too. The room stays about 76 or so now. I may struggle with the iso until it cools off again. The picture of the 1/2g is on right and my clean on left. The green is slight but definitely there.

Hi Daniel, how is the iso doing?

~~~~~~~~~~~~~
Ken