My T.iso journey.....

My T.iso journey.....

Postby tank1980 » Sat Jan 29, 2011 10:09 pm

Hello all,

I have started a T.iso culture and wanted to share my journey as well as get feed back from others in hope that it will help all with this touchy phytoplankton.

I started with two liqiud vials from (FAF) on Jan 28, 2011.

I used 200ml of boiled water (to sterilize) with a salinity of 1.023

I bleached a 16fl bottle and rinsed with boiling DI water.

After the temp of the culture water and vials were at 23C I combined the two along with 0.02ml of silicate and 2 drops of Micro algae grow.

Using a 15w T8 plant light I have been doing 18 hours on 6 hours off keeping the temp at 23C.

As of now it is a nice golden brown color, I have been swirling it around before I turn the light on along with keeping a nice boil of air to keep it suspended. I will keep posted on the progress as I go on. I plan to add 300ml once the color gotten darker in 5 to 7 days and add 300ml more each time I get to a desired density needed.

Any feedback would be great and thanks for the time as well the input from your experience.....

Pete
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Re: My T.iso journey.....

Postby johnrt » Sun Jan 30, 2011 6:52 am

Please do keep us up to date. T-Iso has given many of us headaches.

I thought T-Iso liked it cool. 23 C seems a bit warm, but then, I have never maintained a THRIVING T-Iso culture. They just tick over and threaten to crash from time to time. Infuriating.

How are you judging culture density? Additionally, when you get there, what rate of harvest are you/will you be, using?

Thank you,
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Re: My T.iso journey.....

Postby jolson » Sun Jan 30, 2011 7:04 am

why the silicate I didnt think that T-Iso needed that and what product did you use. How did you measure out .02 ml that would be less then a single drop I think
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Re: My T.iso journey.....

Postby JimWelsh » Sun Jan 30, 2011 12:27 pm

The silicate is not necessary with T-Iso. You might want to try running it a little cooler, if you can.
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Re: My T.iso journey.....

Postby tank1980 » Sun Jan 30, 2011 12:28 pm

I have it at the temp the room gets/stays and have just recorded all the cultures to keep the same temp consistency so 23C was what I could do with ot further actions. As far as density goes I was (told) by a few that when it gets a dark chocolate color to split or increase water volume. I need to get a density measurer but figured since it is started at such a small volume my eyes will have to be the judge for now. I was planning to try different aproaches when I harvest such as restart a few at the same way I started as well as save some for backup until I an get a grip on this stuff. From what I have read it is important to add silcate to reach proper desity and I got that from (FAF) I used a syringe from a test kit cause the tip allows me to pull such a small amount (and yes it was like half a drop :shock: ) I am still very new at this and only have cultured nano. I have wanted to give breeding a try and have been told this is an important phyto to use in order to raise A.tonsa...... I will keep you guys posted of my achievements as well as mistakes.
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Re: My T.iso journey.....

Postby tank1980 » Sun Jan 30, 2011 12:31 pm

JimWelsh wrote:The silicate is not necessary with T-Iso. You might want to try running it a little cooler, if you can.


Ok thanks perhaps I can move the culture to a cool closet what would be a good temp? I was reading that aroud 20C-25C was fine. Also with the silicate do you think chances ar better without than with?
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Re: My T.iso journey.....

Postby JimWelsh » Sun Jan 30, 2011 12:38 pm

I personally culture C-Iso (a different strain) at 23C +/- 1C with great success, but much anecdotal evidence supports staying more around 20-21C if you can. The silicates tend to precipitate out of the media if not consumed, and that is why they have been removed from the forumla for Micro Algae Grow (which is Guillard's F/2 minus silicates). You only need to add them back in if you are growing algae that needs them, like many diatoms do. Isochrysis does not need silicates.
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Re: My T.iso journey.....

Postby tank1980 » Sun Jan 30, 2011 12:50 pm

JimWelsh wrote:I personally culture C-Iso (a different strain) at 23C +/- 1C with great success, but much anecdotal evidence supports staying more around 20-21C if you can. The silicates tend to precipitate out of the media if not consumed, and that is why they have been removed from the forumla for Micro Algae Grow (which is Guillard's F/2 minus silicates). You only need to add them back in if you are growing algae that needs them, like many diatoms do. Isochrysis does not need silicates.



Ok thank you......
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Re: My T.iso journey.....

Postby natureAddict » Mon Jan 31, 2011 4:44 am

just curious, how do u keep 23C during the summer months.
thanks
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Re: My T.iso journey.....

Postby sunny d polyp » Mon Jan 31, 2011 1:35 pm

I've been culturing Tahitian isocrysis galbana for 1 1/2 years successfully and my temps range from 76F to 82F depending on the time of year here in Fl. (US) so I'm some what perplexed on those saying to keep it below 73F, is galabana different from the other T iso you are talking about? I even had one of my iso cultures crash and instead of wasting it I dumped 2 or 4 hundred thousand roti's (plicitilis) and within 3 days turned back to a light golden brown sieved it 4 or 5 times and saved the culture, I culture nano for my roti's but enrich with the iso but this told me that since I assume it was nano that infiltrated my iso and the rots went for the nano first (maybe size factor?) and left the iso alone, my question for some of the guru's here is if the rots didn't have anything but the iso would/could they then consume it eventually, I know they don't seem to want the tet, I dump that in my copes (tigriopus) and fuges.
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Re: My T.iso journey.....

Postby JimWelsh » Mon Jan 31, 2011 1:56 pm

Intersting news, sunny! I've never had a problem, personally, with the warmer temps, but I've heard others report anecdotally the requirement for T-Iso to be grown cooler.

I am able to keep my cultures at 23C in the summer because they are in my basement, and the ambient temperature there is pretty stable. It is pretty well insulated. The Air Conditioning keeps the room cool in the summer, and the other tanks (lights) keep the room warm in the winter.
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Re: My T.iso journey.....

Postby sunny d polyp » Tue Feb 01, 2011 8:47 am

Thanks Jim.
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Re: My T.iso journey.....

Postby tank1980 » Sun Feb 06, 2011 3:12 am

Ok so today will be day 9 from starting my culture and it has gotten to a nice dark golden brown color. I plan on taking 25ml and starting another culture as I did from the startusing 200ml of new water. With the rest I will add 250ml of water to the going culture to increase the amount. This will give me two cultures and I hope that they keep doing as well!!!!

One question is, can I by anyway store some as a back up?

Thanks, Pete
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Re: My T.iso journey.....

Postby sunny d polyp » Mon Feb 07, 2011 8:58 am

I keep a small 500ml flask with about 300ml and split it every 14 days (double the normal time) and wait for it to be at it's darkest because I seem to have much better luck allowing it to be very dark and seems to stabilize it if there is a such thing with iso.

I been toying with freezing it for a few months and seeing whether I can bring it back, anyone ever try that?

I use to keep petry iso dishes handy but I have never had any luck with restarting a culture with them, I've always had to restart it with liquid cultures.
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Re: My T.iso journey.....

Postby JimWelsh » Mon Feb 07, 2011 11:47 pm

You will find a thread in the "Research Projects" forum about freezing, thawing, and re-starting Isochrysis. Anecdotal reports say it is possible. Conventional wisdom says it is not possible. Nobody has been able yet to provide a reproducable method for freezing and then restarting Isochrysis.

I have restarted Isochrysis from a culture held for one year in a test tube at room temperature and ambient light levels with no aeration.
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Re: My T.iso journey.....

Postby sunny d polyp » Tue Feb 08, 2011 8:48 am

I found this link the other day and it addresses freezing a culture in it's exponential phase but adds methanol and freezes it with liquid nitrogen warms at room temp and ends up with a viable culture, was the year old test tube you restarted an axenic culture? It is getting difficult to find anything except uniagal cultures anymore, Seahorse Source use to sell them but now I can only find 3 here in the states and they are a bit expensive. I don't really worry too much about pure nano or tet because they are cultured next to each other and end up with a mix anyway, I restart them with petry dishes from FAF every 3 months.

Jim, what is the longest you have been able to keep a culture of Iso going with out restarting it?


http://web.biosci.utexas.edu/utex/protocols.aspx
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Re: My T.iso journey.....

Postby JimWelsh » Tue Feb 08, 2011 12:17 pm

Allow me to clarify my comment: I meant a freezing/thawing protocol that is practical for the hobbyist!

My year-old test tube was not axenic. It was just a sample taken from a dense culture. The longest I've cultured T-Iso without restarting is about 10-11 months or so.
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Re: My T.iso journey.....

Postby sunny d polyp » Tue Apr 05, 2011 1:08 pm

Hey Jim I just found this thread again, I started another culture of iso I bought from FAF (250ml's) and I split them into (2) 500ml flasks, one did very well and now have a liter but the other turned a pale pink and I almost tossed it but instead I sieved it through a 10 micron twice and set it back in the culture station and within a couple of days I noticed it turning to a pale yellow and now it's that beautiful gold again, the pink was something I have never experienced before, I assume since it is a golden brown that I have iso but I'm a little suspicious so I'm keeping it separated for now, have you ever had your iso turn pink?
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Re: My T.iso journey.....

Postby JimWelsh » Wed Apr 06, 2011 2:21 am

I haven't ever seen pink Iso. When I do have Iso crashes, they usually turn green or perhaps a greenish-yellow. Congrats on the save! If it goes a nice dark chocolate brown on you, you can probably be pretty sure about the culture.
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