Culture Journal, Species: Tetraselmis chuii

General
Species:  Tetraselmis chuii
Species description:  Single cell with 4 equal flagella
Culture source (link if possible):  CSIRO Australian National Algae Culture Collection http://www.marine.csiro.a...recid=34955&-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
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 slit as follows, 500ml to two or three new flasks or soda bottles, remainder to fridge to feed as required.  Split will occur depending on dark green 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 Four separate cultures established with 4 x 65ml starter cultures
04/01/2012 2 x 400ml cultures and 1 x 125ml culture provided to another person.
04/15/2012 split 2
04/27/2012 split 3 
 
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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

 
After 3 days (1 day behind main culture)

 
All cultures look to be growing well.  I am planning on doubling the culture water tomorrow.

The Tet is growing well.
 
I am getting a slightly better growth (darker) from the 2 part solution that the FAF micro algae grow, but not significant.
All four are growing well.
 
The two cultures with FAF micro algae grow, each had 400ml removed and 1500ml of additional culture media added to a total of 2000ml.
The single culture with 2 part media had approx 1100ml added to a total of 2000ml.
 
The small flask has and the two 400ml starters have gone to another person.
 
I have approx 6L of light/medium density algae after only a week.
This is growing really well.

The Tet is now at the stage where I think I need to split it into additional cultures.  How dark should I let it get?
 

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/rotifers.
 
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.

I split all of my Tet last night. It was all very dark green.   In all cases (8 containers) I removed 2/3 of the algae 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.

How do you store TET ?
 
Mine are always settling on the bottom and loose their motility...

I just put it in sandwich bags or 2l coke bottles and then in to the fridge.
It does settle a bit, but I feed this to Artemia and it seems to be ok.

It's really just so I don't dump it. The longest so far has been 2 days in the fridge.

Image for continuation report