Culture medium for algae and other critters

[Pat Thielen]

Hello!

Does anyone here have recipes / techniques for making or buying culture medium for algae and various protozoans? I recently bought several cultures and I realized that I know nothing of culturing them. It may well be too late as the cultures will probably collapse soon, but for future reference it would be most useful. Some of the culturing techniques / advice I would be most interested in at the moment would be:

Various algae types, such Spirogyra, Volvox and Euglena. I just ordered 1000mm of Volvox culture medium from Frey Scientific but I would still be interested in any ideas some of you would know. I have cultures of other types of algae but I don't know offhand which ones they are (I'm at work now and so I don't have them in front of me).

Protozoans such as Paramecia, Vorticella, Stentor, Amoeba, and others. I know there are many species; is there one medium that works well for most? I'm doing a hay infusion right now but I'm not sure if that would be good for everything. I suspect the Stentor and Amoeba may need something different.

I know this is very short notice -- I should have prepared this stuff long in advance. So, if these cultures all crash and burn I'll take it as a learning experience and start over. I'm also plotting on setting up an artificial pond of sorts to grow a community aquarium of microbes; that one is still a work in progress. In any case, I would greatly appreciate any advice any of you would be willing to provide.

Thanks!

[75RR]

Came up with a couple of pdf on this: viewtopic.php?f=15&p=40144#p40144

I have never tried this, I tend to let my samples evolve on their own.
Bought cultures are different of course. Let us know how it goes.

[Pat Thielen]

Came up with a couple of pdf on this: viewtopic.php?f=15&p=40144#p40144

I have never tried this, I tend to let my samples evolve on their own.
Bought cultures are different of course. Let us know how it goes.

Thanks! I'll give these a look.

[billbillt]

Came up with a couple of pdf on this: viewtopic.php?f=15&p=40144#p40144

I have never tried this, I tend to let my samples evolve on their own.
Bought cultures are different of course. Let us know how it goes.

Algae seems to be self-sustaining.. I have a sample in a small jar sitting where it can get sunlight with the label on it marked: "Bucket Water 11-09-2012".. It is still living... The sample came from a bucket set out to catch rainwater as it runs off of the roof..

BillT

[zzffnn]

Plant like algae mainly needs light and heat. You may try (I am not positive) instant flower food in those tiny bags, but be very careful not to add too much. They may need some nitrogen and phosphorus, but too much will kill them.

Exactly what and how much is science.

Animal like protist, such as flagellates (Euglena) and ciliates need something different. I would suggest:
1) carbohydrates (rice starch);
2) yeast abstracts (that provides amino acids and some B vitamins);

3) heat-inactivated bovine serum (that provides lipids, growth hormones and proteins). This is what researchers routinely use for culturing eukaryotic cells. Maybe useful but expensive. Here is a crazy idea: get some animal (or your own) blood, spin down/remove red cells, heat inactivate it (look up what temperature) and use it that way. Skip if in doubt

4) diluted fish food/pellets

Be very careful with other ingredients such as vitamins/salt/iron/minerals. Too much will kill. Skip if in doubt.

The ATCC guide linked by 75RR is not very specific for pond protists and only gives some hints of basic requirements. Their protocol is only specific with pathogenic eukaryotes (parasites), because researchers routinely culture those for scientific projects and people have devoted time to develop specific culture media.

[einman]

An excellent guide is :

Culture Methods for Invertebrate Animals by Frank Lutz, Paul Welch Paul S . Schott

Covers nearly every class..

[carlh6902]

This may be of some use...

http://static.fishersci.com/cmsassets/d ... Manual.pdf

Carl

[charlie g]

Hi Pat and all...happy, healthy new year. Think about a simple aquarium 15 gallon tan k (low cost...even lower cost if purchased at a 'flea market/ garrge sale'.

Plunk an air-stone bubble source in the little tank...plunk in three or four low cost 'feeder fish', or guppies from a pet-shop...feed these few fish only 2X/week....this your microcosm for algae culture. charlie guevara

[Pat Thielen]

I made a hay infusion almost a week ago but so far I haven't seen large numbers of microbes appearing. There are some, but not very many. I started up another one, this time I boiled the hay for around 15 minutes which supposedly helps it break down a bit and encourages growth of bacteria and other microbes. I just started the last one today, so I'll have to see how it goes. I also added a drop of milk to it to hopefully get things started (I think that was part of the recipe I found on the Microbe Hunter website.

I hope the Volvox culture medium doesn't freeze when it arrives...

[75RR]

... this time I boiled the hay for around 15 minutes which supposedly helps it break down a bit and encourages growth of bacteria and other microbes.

Never done one but I thought one had to seed the resulting hay infusion with the desired protists.

[JimT]

Pat, my experience;

During the warm months I collect a few jars of pond water. After a while there is not much life left so I add salad greens to the jars. Within a few days the jars are teaming with protists.

If Minnesota has a thaw any time soon collect some pond water and "nurture" it.

Otherwise, over water your house plants and collect the overflow in the tray under the pot. I just observed some and was surprised at the amount of life there was.

Let us know how you fare.

JimT

[Pat Thielen]

... this time I boiled the hay for around 15 minutes which supposedly helps it break down a bit and encourages growth of bacteria and other microbes.

Never done one but I thought one had to seed the resulting hay infusion with the desired protists.

What happens is rather strange in a way (and makes the idea of "spontaneous generation" quite believable). The first thing that happens is bacteria appear and feed on the plant material (in this case hay). And then, a species of protist appears that feed on the bacteria. I don't recall the species, but this has happened both times I've made a hay infusion and I strongly suspect it will be the same for the third. Anyway, the larger cilates will feed on those protists that appear along with the bacteria.

This will eventually become a paramecium culture medium and should also be able to take care of a few other species.

[Pat Thielen]

Well, I tried to cut out the middlemen and made my own Volvox culture medium. My thought process was actually very simplistic: Volvox are basically algae and so maybe something that would feed plants would also feed Volvox. So, I took 1/4 teaspoon of Gerten's plant food and dissolved it in 500mm of distilled water. I then put some in a small jar and introduced Volvox to it. That was on Wednesday and so far the Volvox seem happy enough and don't appear to be dying off. This seems to be a good sign -- I was worried that the distilled water may cause trouble with osmosis (this can cause protists to explode) and that the mix would be too strong. So, I'll observe them over the next few days and see how they do. If it works, it sure beats paying $20.00+ for a liter of Volvox culture.

I'll let you all know how it goes.

[einman]

... this time I boiled the hay for around 15 minutes which supposedly helps it break down a bit and encourages growth of bacteria and other microbes.

Never done one but I thought one had to seed the resulting hay infusion with the desired protists.

I have had a Hay infusion going since October of 2015. A lot of copepods..

[einman]

My culture food of choice is Tetramin fish flakes.

I feed it to my crickets, mealworms, Collembola and teh hay infusion. I also fed it to the my fish..well it is fish food..LOL

[Pat Thielen]

My experiment with the plant food culture medium was a bust. The Volvox seemed happy enough for a time, but eventually died off and all that's left (by direct observation) is blue-green algae. I haven't looked at it under the microscope yet, but will hopefully soon. I wonder if I should have boiled the water first, or if perhaps the plant food content was too strong. Maybe the Volvox need something in their cultures I don't know about. It would be very cool to be able to keep these guys going. I didn't think it would be all that difficult as they are plants.

Anyway, if any of you may have some insight into this I would greatly appreciate it.

[rabitt]

Pat,
try this site, http://www.microscope-microscope.org/ap ... otozoa.htm

I have a few more sites if you need them.

Rich B

[kinase]

Oh I wish I saw this sooner... My lab is an algae zoo, complete with various kinds of media (many of which are highly similar).

Is this fresh or salt water we're talking? There's lots of recipes out there, but you kind of need a small chemistry lab to make them. You can probably boil solid fertilizer in water and use dilute versions of that. Too much and the bacteria will take over. https://ncma.bigelow.org/ is where you want to look. If you bought a media kit, you can make many liters of media, https://ncma.bigelow.org/products/growt ... edia-kits/

A little pond is going to be pretty tough, you throw enough stuff in there and they're going to start killing each other until you reach a balance. Cyanobacteria and regular bacteria grow really fast if you add too much nitrogen or phosphorus, or too much carbon.