Culture Journal, Species: Isocrysis galbana (failed)

General
Species:  Isocrysis galbana
Species description:  Golden Brown motile alga
Culture source (link if possible): Florida aqua farms http://florida-aqua-farms...;product=LIVE_CULTURES
If algae, CCMP # (Optional): 
http://ccmp.bigelow.edu/
Culture Establishment Date:  12/3/2011 (3rd December 2011)
Continuation Date: 

Culturing Vessel Details
Salinity:  1.025
Temperature:  not measured 
pH:  not measured

Vessel description:  2L cola bottle with airline.
Lighting description:  Algae Cupboard with three vertical double fluro daylight bulbs (1 bulb of each bank is disconnected due to excess heat).
Lighting cycle:  16 On / 8 off
Aeration description: Rapid aeration

Methodologies
Split methodology:

Culture medium description: 
Sterile Artificial Salt water with 1.4ml per L of Florida Aqua Farms Micro Algae Grow

Cell count:
 (if known)

Reference links:  

Additional Information
(No Pictures or Videos in the Section Please)
Notes: 



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Starter was a FAF culture disk. I am aware that people have had difficulty with these in the past (lots of difficulty). Unfortunatly it was my only choice to attempt an ISO starter culture. The process was as per the FAF instructions.
 
I had two culture disks, so I used culture water (1.02 artificial aged salt water) that had been microwaved for 7 minutes and then allowed to return to room temprature.
From this culture water, enough was removed to cover each of the culture disks.
These were then covered and placed under a light for 24 hours.
The culture's where then removed from the disks with a cotten tip. The algae was added to a 2l container with 500ml of culture water and 5 drops of FAF algae nutrient (approx 0.7ml).
 
The bottles have been placed in my Algae cupboard at a distance of about 15cm from the fluro globes.
 

 
 

As of today, the algae appear to be keeping the water tinted (yellow rather than brown) and some are settling on the bottom of the containers.
Twice a day I move the ridged air line around and return the algae to suspension.

I may be mistaken but I'm pretty sure microwaving SW is not a good idea...

http://www.fishtalpropagations.com/#!home/mainPage
"Making captive breeding easier."

I microwave all of my phyto culture water until it is over 185F / 85C.  Microwaving is recommended as a sterilization method for phyto cultures in the Phytoplankton Culture Manual - 6th Edition by Hoff & Snell, page 39.  I've even recently started adding my F/2 *before* microwaving, and have had better success preventing contamination as a result.  I do see some precipitate after microwaving, but my phyto cultures are still going strong!
 
I think the main concern is to prevent actual boiling, and the subsequent precipitation of substances out of solution.  I know that Martin Moe has mentioned microwaving to 185F, waiting 24 hours, then microwaving to 185F again, and letting the media cool, to ensure sterile phyto media (I can't find the reference right now).
 

I did find this:  http://www.sciencedirect..../pii/0022098188900639 which is referenced in this document (among others):  http://nsgl.gso.uri.edu/flsgp/flsgph07002.pdf

I followed some instructions from FAF that said to use sterilized water, then suggested using a microwave.

It's seemed reasonable.

Thanks for the info though, it's useful.

Microwaving as described above will sterilize saltwater as well as freshwater, and it will sterilize a soda bottle if that is what you are using. Unfortunately, putting hot water in a soda bottle will melt it.  I use bleach for that reason, because I use soda bottles.
 
Also, I've heard of many hobbyists using FAF disks to start Isochrysis, but I've never heard of any that had a single success.  I hope you will be the first. But don't be discouraged if you are not.  Algagen, SEahorse Source, and many others sell liquid starter cultures.  Not that I've had any luck lately, but I think its my 6 year old food.  THAT, Micro-algae Grow, you should get from FAF.  Also, their nanochloropsis disks work very well.
 

Quick question, how did you source your disks from FAF into Australia?
cheers
Captain

I ordered it from their web site.

Thanks for that. I would have thought there would be problems getting live cultures into Australia from abroad with quarintine laws, aquis etc.
cheers
Captain

Yeah, I was half expecting it to be confiscated.

I filled out the customs declaration with the full species and made it clear that it was algae.

The box was opened by customs or quarantine (I don't remember which one) because a piece of paper was in the box saying so, but it came through.

At the end of the day, I declared it correctly and it was allowed in so that was good.

Wow. I might have to give it a go myself in the new year after holidays. I want to start up some additional algae cultures apart from nanno. How did you sort the paperwork out, does Florida Aqua Farms send you the forms or do you have to contact customs yourself?
cheers
Craig

OThe customs stuff was done with the freight forwarder (shipito I think).

Unless you really want live you can buy the Reeds stuff from proaquarun qld, or nano, tet and a few others (mixes) from reefculture again in qld.

These are both much easier (and cheaper in the long term) than growing it. It becomes, take out of fridge/freezer, defrost it and use.

I'm only growing the iso to have a fall back if the reeds iso concentrait does not work well enough for the Copepods I'm getting. I hope to be able to discontinue this culture.

Ok thanks mate. I have purchased stuff from proaqua before I think it was rotifer diet. I just get better results when using live cultures so that's why I'm thinking of adding additional types of cultures. Out of interest what copepods are you getting? Have tried culturing some pods harvested from my refugium without success, not sure of the variety they were though.
cheers
Captain

At this stage Im getting Gladioferens imparipes plus some others from the the School of Marine and Tropical Biology once we sort out which ones.
 
At this stage, the hold up is getting ISO that will grow in enough volume to support them.

This appears to be a complete fail.  All I have at this stage if filaments of some type of algae on the bottom of the bottles and clear water.
Im letting it go for a few more weeks, but it looks done to me.

Freakin' Iso.... Sorry to hear of the crash, but it's all too common.

--Andy, the bucket man.
"Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886