Culture Journal, Species: T.Isochrysis sp

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CaptCrash
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Culture Journal, Species: T.Isochrysis sp - Thursday, March 22, 2012 11:22 AM
Culturing Journal DataSheet
This first post should be updated regularly to include new information as events take place or changes are made to your system

General
Species:  Isochrysis sp
Species description:  Cells with 2 equal flagella and short haptonema
Culture source (link if possible): CSIRO Australian National Algae Culture Collection http://www.marine.csiro.a...recid=35112&-find=
If algae, CCMP # (Optional): 
http://ccmp.bigelow.edu/
Culture Establishment Date:  03/27/2012
Continuation Date:  06/25/2012

Culturing Vessel Details
Salinity:  1.015
Temperature:    Not measured (Fans turn on at 20c to cool algae cupboard)
pH:  Not measured
Water Details: Artificial Salt with RO/DI water.

Vessel description:  2000ml Erlenmeyer Flask/Conical Flask and plastic soda bottles
Lighting description:  CF Light, 3 x double bulb, 37W fixtures.  4 x 5000k bulbs installed (2 spares)
Lighting cycle:  16 hours per day, 7am to 11pm
Aeration description: Heavy Aeration, adjusted to rotate culture approx 1 per second.

Methodologies
Split methodology:  Cultures are split, 500ml to two or three new soda bottles, 500ml to a new clean flask.  Split depends on dark colour/density of culture.

Culture medium description: 
RO/DI water with Artificial Salt to 1.015.  Micro Algae Grow┬«, Florida Aqua Farms Inc added at a rate of 0.4ml / 1000ml salt water.
Media is microwaved in an 1100W microwave for 17.5 minutes per 2.5L batch.  Batch is allowed to cool to room temperature before use.

Cell count:
 (if known)

Reference links:  

Additional Information
(No Pictures or Videos in the Section Please)
Notes: 
03/21/2012 125ml of sterile source algae was ordered from CSIRO (order was for 250ml and was shared with another person).
03/27/2012 Cultures established with 4 x 65ml starters
04/01/2012 2 x 400ml starters plus 1 x 125ml starter provided to another person.
04/15/2012 split 2
04/27/2012 split 3

You will be required to provide photographic evidence and as much detail as possible about your project in this thread.
If your thread does not contain detailed enough photos  and information the MBI Council will not be able to approve your reports.

<message edited by CaptCrash on Monday, June 25, 2012 4:29 AM>

CaptCrash
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Re:Culture Journal, Species: T.Isochrysis sp - Saturday, March 31, 2012 11:45 AM
Pictures of Algae as supplied by CSIRO in Australia.  They provide pure samples and it's a much more significantly presented to the last liquid culture I received as a starter (in an empty coke bottle).
 
The box arrived

 
Including this warning

 
Inner Box

 
Packaging (cotton wool)

 
Packaging (foam beans, more cotton wool, each flask individually wrapped)

 
The flasks

 
65ml was added to 1000ml of media in each 2000ml flask.

 
The flasks were sealed with sterile cotton wool.
 
Each of the flasks and tubing were microwaved for 7 minutes with 1000ml of water in an enclosed container in an attempt to ensure that they were as sterile as possible.
 
A total of four containers have been used.
2 x 2000ml flasks.  Two of these flasks contained FAF Micro Algae Grow at 0.4ml/1L, 65ml starter with 1000ml media added.
1 x 2000ml flask with a two part F/2 solution I have had for about 18 months, 65ml starter with 1000ml media added.  This was added at 1ml each of parts A & B per 1L.  I am unaware of the "brand" of this F/2, But I have approx 300ml of each of the 2 parts and wanted to see if it was still good.
1 x 250ml flask water was added that contained FAF Micro Algae Grow at 0.4ml/1L, 65ml starter with 65ml media added.  This is the backup sample.  It is being kept separately to the main cultures under a CF globe and has an air line bubbling in it.
 
After 4 days

The flasks look slightly less brown/golden than this image shows
 
After 3 days (1 day behind main culture)

The T-ISO actually looks slightly more brown than this image shows.
 
Two of the three 2000ml flask cultures look to be doing well, one does not seem to be going well.
The small flask is going ok, it was looking like the only one growing until today.
<message edited by CaptCrash on Tuesday, April 3, 2012 8:16 AM>

CaptCrash
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Re:Culture Journal, Species: T.Isochrysis sp - Sunday, April 1, 2012 7:22 AM
The two cultures in glass flasks that are progressing very well and are golden brown today.
They each had 400ml removed from them to supply to another person.
The remaining 500ml had 1500ml of culture media added.
These are one each of of the FAF media and the 2 part media.
 
The culture in the glass flask that is not progressing as well as the other two, is showing some improvement and was also topped up to 2000ml total (adding about 1200ml of FAF culture media).
The small flask was also doing well, but has gone to its new owner.
 
I now have approx 4L of T-ISO that is light, but appears to be of a good colour.

CaptCrash
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Re:Culture Journal, Species: T.Isochrysis sp - Tuesday, April 3, 2012 3:47 AM
Cultures are doing very well, this picture does not quite show how golden the algae is, it seems to be much thicker/darker in person with quite a glow.
 

<message edited by CaptCrash on Tuesday, April 3, 2012 8:19 AM>

CaptCrash
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Re:Culture Journal, Species: T.Isochrysis sp - Saturday, April 14, 2012 11:16 PM
A split of one flask was performed on 04/11/2012, taking 800ml from the conical flask and adding 400ml to two 2L soda bottles.  All three containers were then filled to 2L with new media.  Loss from the original 2000ml is due to evaporation and use for fry.
 
A split of the two remaining flasks was performed on 04/15/2012, taking 800ml from the conical flask and adding 400ml to two 2L soda bottles.  All three containers were then filled to 2L with new media.  Loss from the original 2000ml is due to evaporation and use for fry/rotifers.

JimWelsh
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Re:Culture Journal, Species: T.Isochrysis sp - Sunday, April 15, 2012 7:47 AM
Great news!  Can we see some pictures of some nice, dark, chocolate brown cultures yet?

CaptCrash
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Re:Culture Journal, Species: T.Isochrysis sp - Sunday, April 15, 2012 9:53 AM
Quote Originally Posted by JimWelsh

Great news!  Can we see some pictures of some nice, dark, chocolate brown cultures yet?

JimWelsh, Lucky you should ask.
 
I ran out of culture medium whilst doing the last flask and had to make more and let it cool.
You posting this reminded me that I should take a picture, so here they are (I had already taken the first split amount out, so this is a bit low).
 
Picture with side lighting and flash (a little darker than to the eye)

 
with back lighting and flash (a bit more of a glow than to the eye)

 
The real colour before the split is a bit lighter than the first picture.  Its not really got a golden colour at this stage, the ones I have split look like they will be the brown/golden colour in a day or two.

JimWelsh
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Re:Culture Journal, Species: T.Isochrysis sp - Sunday, April 15, 2012 11:59 AM
That's the stuff!  Looks great.

Zooid
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Re:Culture Journal, Species: T.Isochrysis sp - Sunday, April 15, 2012 8:17 PM
Congrats....I've never had good experience with T-iso.  Very impressive

CaptCrash
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Re:Culture Journal, Species: T.Isochrysis sp - Friday, April 27, 2012 3:00 AM
I split all of my ISO again last night.  I probably left some of it a little long and it was very dark.
Its growing really well and touch wood, its growing well.
In all cases (6 containers) I removed 2/3 of the ISO and used it to feed rotifers/artemia, then topped up with new media, the remainder was stored in the fridge for use in the next few days.

CaptCrash
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Re:Culture Journal, Species: T.Isochrysis sp - Monday, June 25, 2012 2:17 AM
Image for continuation report


nrbelk
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Re:Culture Journal, Species: T.Isochrysis sp - Tuesday, February 12, 2013 12:51 PM
Any updates?

CaptCrash
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Re:Culture Journal, Species: T.Isochrysis sp - Tuesday, February 12, 2013 7:22 PM
Not really.

I find t-Iso really annoying, it is allways in the verge if failing.
I had to start a new cloture about 4 weeks ago as the heat here in Perth finally did it in.
It has been getting to mid 40c for days since Christmas.

To be fair I also lost the tet and nanno cultures around the same time.
They are all just starting to come back now.