My first algae culture experiment

[Crater Eddie]

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This is my first attempt to culture some algae. The left bottle is from a pond that I have been sampling since high school biology, 30 some odd years ago. I intentionally avoided collecting any visible clumps of algae. The bottle on the right is from my dad's pond, this time including a big clump of algae. I'll set up another bottle using some dried up flakes of algae from my birdbath, but I need a manifold for the air hoses so I can run more than two bottles, and some valves to mediate the bubble stream.
I'll need something to feed the algae, I wonder if I can use a little African Violet fertilizer, or maybe a touch of Miracle Grow?

[lukem321]

One suggestion: I read that air stones can inhibit algae growth and that bigger bubbles are better. Other than that, looks good

[Crater Eddie]

Oh, that's good to know, thanks!

[Crater Eddie]

The thing that irks me is that about a month ago I bleached all my water storage bottles that I use for my carnivorous plants, because they were getting... algae buildup.

[Crater Eddie]

I removed the air stones and attached a pipette to each tube instead... that allows me to still get the end down near the bottom of the bottle and still produces fairly large bubbles instead of the maelstrom produced by the stones. Also plugged the air pump into a timer and set it to run for an hour at a time a few times throughout the day and night instead of just always on or always off. That way they still get some air, but still have time to "relax" in case that is helpful. When I find some valves I'll turn down the bubbles to a trickle to reduce the turbulence, hopefully minimizing trauma to the organisms. Now if I could find some #2 rubber stoppers and a borer I could do it up properly.

[Crater Eddie]

Searching online for information on culturing algae results mostly in growing algae for making biodiesel, and spirulina for nutritional supplementation. Interesting subjects, but not really what I was looking for. Then I found the key: instead search for "culturing phytoplankton". This turns up tons of information and even Youtube videos by and for aquarium hobbyists for growing fresh live food for fish. Most of the examples are saltwater cultures, but the principles apply to fresh water organisms as well. Lots of good practical information.
Thanks to lukem321 for sparking my interest in this.

[75RR]

I was wondering if one was going for Pediastrum for example, then several smaller bottles with algae from several sources might provide more of a chance of success.

[Crater Eddie]

Quite possibly. I think it might take a lot of effort to isolate a pure culture from a wild source, but it should be do-able given time and determination.
Of course I am just testing method here to see what will grow, mainly for entertainment. There are sources online that sell pure cultures of just about anything you could want, but this is more fun.
CE

[gekko]

Looks like a serious effort . I hope that we'll soon see some wonderful images of whatever will grow in there. Thanks for the pictures.

[lukem321]

75rr: That's pretty similar to what's called a diltuton. For a dilution you basically just take a drop of algae-dense pond water and put it in a test tube full of clean water. Then take a drop of from that test tube and add it to another. If you do this enough times, there's a chance you get a few cells of just one species in a test tube. The problem with this is that it is very uncertain, will probably take a lot of tries, and will take months for the few cells to grow into a culture.

[Crater Eddie]

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The bottom shelf on my plant rack doesn't get all that much light, so I added a 100w equivalent CFL on it's own timer. We'll see if it's enough.

[gekko]

Nice setup (except I'm a bit nervous about the lamp being so close to the water...).

[Crater Eddie]

Thanks Gekko. Thanks for your concern, but there really isn't anything to worry about. The lamp is no more at risk than a lamp in a fixture above an aquarium. In this case, it is separated from the water by the thick glass of the wine bottles rather than the flimsy plastic of the aquarium lamp fixture. There is no spray coming from the bottle mouths either, and I have the base of the lamp well insulated.

The plankton culturing folks use a fancy fertilizer specially made for algae, though if you dig enough you can find many that use Miracle Grow liquid plant food and say it works just as well. With this in mind, I added 10 - 12 drops of Miracle Grow liquid African Violet food to each bottle, just because that's what I happened to have on hand. The dense clump of algae in the "dad's pond" bottle is now a noticeably darker green, so I must assume that it has done some good. Some of the other bottles are starting to get a thin film of algae sitting on the bottom, so this is good too. After it gets a bit thicker, I might move the bubblers around a bit to try and break up the film and keep it suspended. Maybe. Or maybe not.
Oh, and that bottle on the right has several flakes of (formerly dried) algae from my birdbath. I'm hoping that it will grow some nice haematococcus. I have a nice culture growing in a petri dish, but it will be fun to see if this works too.

[gekko]

Thanks Gekko. Thanks for your concern, but there really isn't anything to worry about. The lamp is no more at risk than a lamp in a fixture above an aquarium. In this case, it is separated from the water by the thick glass of the wine bottles rather than the flimsy plastic of the aquarium lamp fixture. There is no spray coming from the bottle mouths either, and I have the base of the lamp well insulated.

Sorry for my paranoia, but somehow the proximity of a live bulb socket to jars full of aerated water worried me a bit.

Back to the main subject: On the main MicrobeHunter website there are a couple of posts by Oliver (there might be others) that might also be of interest:
http://www.microbehunter.com/growing-algae-in-a-jar/
http://www.microbehunter.com/making-a-s ... ing-algae/

[Crater Eddie]

Thanks for the links, I had not found those.

[Crater Eddie]

After having to make a long hook to pull some filamentous algae out of one of these bottles, I decided I needed something better.
This old pickle jar was the perfect solution... I knew I kept it for a reason. I have always collected jars, especially large ones.





It's a good thing we like pickles.

[75RR]

This old pickle jar was the perfect solution...

Hope this does not mean you are giving up wine!

[JimT]

Can't wait to see photos of the algae. Seems like a lot of work for pond scum and slime
How soon before you have green stuff?

[Crater Eddie]

Hope this does not mean you are giving up wine!

Perish the thought!

[Crater Eddie]

Can't wait to see photos of the algae. Seems like a lot of work for pond scum and slime
How soon before you have green stuff?

Jim, I have posted a few pics from the haematococcus bottle as well as a few from the filamentous algae bottle in other threads, perhaps you missed them.
I checked one of the other bottles last night and found a very very small predominant algae, I haven't identified it yet. If I get any good photos of it I will post.
CE

[Crater Eddie]

I noticed this morning that my haematococcus culture has collapsed. The water is now almost clear with a thick layer of pink at the bottom. Next time I'll try starting a new culture when the red peaks in intensity. Is the layer at the bottom dead algae, or has it just gone dormant? I'll examine a sample this weekend and start a new bottle, either with some of this or some new flakes from the birdbath. I wonder if the jar ran out of nutrients? I didn't feed it after the initial inoculation, maybe I should have.
CE

[gekko]

It might be worth looking at a sample from the pink layer at the bottom to see what it is: might it be a layer very rich in Haematococcus?

[Crater Eddie]

That's what I am hoping Gekko. Surely that is what it is, I just don't know if they are alive, dormant, or dead. I'll get a chance to look later tonight, I hope.

[Crater Eddie]

I finally got a chance to look at the haematococcus bottle tonight. The water isn't quite clear, it does still have a red tint to it, but there is a thick layer of red stuff at the bottom.



This came out in the pipette as a thick sludge, and made a very thick drop on the slide. I had to swirl it around with a needle to even it out and dilute it with a little distilled water so the cover slip would settle. Examination through the microscope revealed a thick soup of haematococcus, mostly completely motionless, with no flagella evident. It took some doing to find a few out away from the huddled masses. They don't look dead, but are completely motionless, so are they dormant? There were a few swimming around, very few.



The most impressive thing about this sample is that the whole slide was practically jam packed full of rotifers.



They were everywhere, and most of them were stuffed with red... apparently haemotococcus remains.



There are a few green algae here and there, and plenty of amoebae.
I think it's time to start a fresh culture.
CE

[gekko]

Nice pictures.

I think it's time to start a fresh culture.

...and also to post more pictures of rotifers, amoebas, and resting-stage Haemotococcus (if not swimming ones) from this critter-rich bottle .

[Crater Eddie]

I decided it was time to start a new Haematococcus bottle. I put it on the back of the shelf, away from the lights, in hopes that the Haematococcus will stay green and motile longer, and not encyst so fast.




Here you see where I scraped my inoculant from the side of the original bottle. I also took several cc's from the sludge on the bottom.





Now we'll see how long it takes to multiply and fill the new bottle.

[gekko]

Interesting project. Meanwhile, I hope you will show us more interesting critters from the other jars.

[Crater Eddie]

I have been keeping an eye on the other bottles, but so far the contents have been pretty boring. A couple of them are starting to get nice and turbid now though, so maybe it's time for another look. Stay tuned, as they used to say.
CE

[Crater Eddie]

By the way, this arrangement of bottles around a single bulb is quite compact, but sitting on the bottom shelf like this makes accessing the bottles in the back somewhat problematic. I am thinking of switching out the CFL for a standard fluorescent fixture so I can just line them up. This would reduce the efficiency of the setup, but would be a heck of a lot more convenient.
Just an observation for anyone who might be considering setting up such an arrangement.
CE

[einman]

Im with Gekko. More pics of the rotifers etc. I plan on culturing living organisms such as the rotifers, paramecium etc. I may have to review your notes on alage culturing just for this reason.