I was doing a project for a class of mine where I was culturing phyto, and I had to do this write up. I'm going to post it up here as a revision of my first post. I haven't redacted it, although I probably should because I wrote it at 12 at night.
anyway, here goes:
Sterile cultures of Nannochloripsis and Chlorella species were obtained. We also obtained a batch of food-grade Nannochloripsis that we used for culturing.
We used clear uncoloured plastic 2 litre bottles of a regular shape that were sterilized and rinsed with deionized water. Two holes were made in the caps, one larger one, centered, for the airline tubing, fitting snugly, to go through, and a smaller one to let air to escape so as to not build up pressure. Airline tubing was strung through the cap with enough length so as to more than reach the bottom of the bottle and enough to reach the gang valve without being taught. The bottles were stripped of labels to allow maximum light penetration.
To create the initial culture water we used 2.0 litres of deionized water minus the initial culture volume. This volume we would mix in proper amounts of marine aquarium salt mixture in a large clean beaker. (Two of the Nannochloripsis cultures are at a specific gravity of 1.025 one culture is at a specific gravity of 1.015 and the Chlorella culture is in fresh water with a specific gravity of 1.000. These values were determined with a refractometer) To the prepared water at the correct specific gravity we added 1 ml of fertilizer. The fertilizer is a Guillards F2 formulation without silicate procured from Florida Aqua Farms. A list of the chemical makeup is attached. The Fertilizer was prepared per the supplier’s instructions.
The pure culture was added to the prepared water and fertilizer in the beaker and using a funnel the culture was poured into the culturing bottle. The bottle was capped off and the airline hose was connected to the gang valve. The airflow thru the tubing into the bottle should always be enough to keep the algae in suspension. The culture bottles are placed at a light source with a 16 - 18 hr on cycle and a 8 - 6 hr off cycle regulated by a timer. The cultures should be left at normal room temperature.
Splitting of cultures can be based on turbidity (culture density) measurements or based on log phase doubling time. A culture of Nannochloripsis in log phase has a doubling time of about 8 days under ideal conditions. Log phase doubling times vary between species of algae. When splitting a culture, its best split it in half and replace the removed volume with prepared water as per the instructions of creating a new culture listed above. It is essential that the same amount of fertilizer is added as when starting a new culture, especially if trying to maintain a culture in log-phase growth.
I hope, this helps someone out who is interested in doing the same. I should take some pictures of my set up I have at home. I'll try and get those up after I'm thru with finals.
(edit: broken up into sections for readability.)
Remember, copper is one of those funny things...a tiny bit is needed for life, a little more kills things. The micro algae grow has just that right tiny bit.
