Best tips for sterile phyto?

BRS

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Hello,

For all of you that run phyto cultures for long periods of time, could you list your steps as to what you do to keep everything sterile. I think my issue was that my bottles of f/2 got contaminated or my salt mix did (that is my main guess). Also, does anyone know what exactly bacteria (or other microbes) do that crash a culture?
 
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Jay Hemdal

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Hello,

For all of you that run phyto cultures for long periods of time, could you list your steps as to what you do to keep everything sterile. I think my issue was that my bottles of f/2 got contaminated or my salt mix did (that is my main guess). Also, does anyone know what exactly bacteria (or other microbes) do that crash a culture?

I've had protozoans crash a lot of my phytoplankton cultures. f2 Gulliard's can get contaminated, so I try to mix that up fresh more frequently, to try to avoid that. Another trick might be to chlorinate and then dechlorinate your culture media prior to inoculation. I try really hard for a "xeric" culture, but contamination still does happen. My "fix" has always been to culture phyto in a number of 2 liter soda bottles, and then, if one crashes, I just dump it, and sub-culture new bottles from the good ones. Note: I only grow phyto in order to culture Brachionus, so it is possible that I avoid long-term issues because the rotifers get to the algae before it truly crashes on me.

Jay
 

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I add 10 ml bleach to a 5 gallon carboy with fresh saltwater and let it stand for at least an hour. Then, under heavy aeration I add the f/2 fertilizer (using syringes that were sterilized with boiling water) and let it mix for at least 10 minutes. Only then I add heat sterilized Sodium Thiosulfate solution to neutralize the bleach and inoculate the water with a clen culture starter (using sterilized syringes).
Also, make sure you put a 0.2μm filter on the airline going into the culture.
 

manolius148

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For me, antibiotics have worked so far when cyano takes over the culture, but it hasn't proved to be a long- term solution. Experts claim that routine checking the culture with a microscope and centrifugation are the best way to separate pure strains but I have no experience in how to proceed. On Amazon, some basic lab centrifuges are offered for less than $100.
 

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For me, antibiotics have worked so far when cyano takes over the culture, but it hasn't proved to be a long- term solution. Experts claim that routine checking the culture with a microscope and centrifugation are the best way to separate pure strains but I have no experience in how to proceed. On Amazon, some basic lab centrifuges are offered for less than $100.
What antibiotics do you use, and where can I get some? Just had a tetraselmis culture turn into cyano. It's becoming my nemisis.
 

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I add 10 ml bleach to a 5 gallon carboy with fresh saltwater and let it stand for at least an hour. Then, under heavy aeration I add the f/2 fertilizer (using syringes that were sterilized with boiling water) and let it mix for at least 10 minutes. Only then I add heat sterilized Sodium Thiosulfate solution to neutralize the bleach and inoculate the water with a clen culture starter (using sterilized syringes).
Also, make sure you put a 0.2μm filter on the airline going into the culture.
I hadn't thought of the f/2, wonder if that's my problem. I was worried bleach would react some way with f/2, so I've been adding f/2 after neutralizing.

I recently started using UV to neutralize the bleach, wonder if that would mess with f/2?
 
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What antibiotics do you use, and where can I get some? Just had a tetraselmis culture turn into cyano. It's becoming my nemisis.


I personally would not use an antibiotic, as we there is a bit of a crisis of people creating superbugs by frequent and improper use of antibiotics for whatever they are doing.
 
BRS

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