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Thread: Paramecium Culture

  1. #1
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    Paramecium Culture

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    Hi Scott,

    Mmmm................now what do you want me to do with this little bottle I just receive 10 minutes ago? You are the 2nd guy after Keehoe to sent me the creepy stuff in the mail. He sent me Walterworm in a smash-up container with worms crawling out about and Phew! What a smell! I think he did it on purpose, you know what? I am going to sent him some fruitflies maggots in the mail.

    Will I kill them if I pour them into a larger container of aged water or boiled water. How do I prepare the oats for feeding? Do I soak them or just throw some into the culture. Do they need light, aeration and do I put a plastic film over the container like your greenwater?

    Anyway thanks Scott for the early X'mas present. Can't you read the fineprints when I did not respond when you insist on senting creepy crawlies to me. Girls or women preferred gifts like soft toys, jewellery...... and cute cute stuff. Not worms or creepy crawlies!!

    ---------------------------------
    Selena

  2. #2
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    Hi Selena,

    Hmmm there is just no pleasing women is there...

    Ok the paramecium are easy just add some boiling water to a take away container, and add some oats about a teaspoon, wheat or uncooked rice will work also. Add them when it is hot and put the lid on the container and allow to cool. Then add some of the starter culture. That is all.

    Be careful to try and keep the lid on as much as possible, since they are pure cultures and will get infected from the air. So if you open it tip the lid partially open rather than taking the whole thing off. They don't like any bright light just normal room light is fine. Just feed with a dropper and occasionally renew the culture as above, maybe once a month.

    If everything works nice your container should turn nice and milky, and you will be able to see them swimming around in there. In the home there is no way to keep it sterile but really just try and make it as nice for the paramecium as possible, then they will become the dominant organism there.

    And I will see what i can do about a better christmas present for you

    I think I must of caught something from Ronnie.... I have started building stuff and playing with silicon and visits to the hardware. .

    Scott.
    Thanks again,
    Scott Douglass

  3. #3
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    Hi Scott,

    How long does it takes before I get a significant quantity to feed the newborn fry? I have several bags of fulminantis eggs that are due for wetting. Fulminantis fry are very tiny and your paramecuim will definitely increase the survival rate.

    And I will see what i can do about a better christmas present for you
    Thank you Scott, when you get everything done up, just a snap shot of the set-up will do.


    -------------------------------------
    Selena

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    Hi Selena,

    They should take about a week to get a proper culture growing. Not Long at all. They are very good for the smaller fry, especially those fry that seem to be left behind. Slower growing.
    You can check your culture with a magnifying glass against the light, you should see all the little dots actually moving around. I would still also use the walter worms and anything else you have, since it is good to give them as much food choice as possible. But definitely it should make a difference with small fry.
    BBS are great but for some fry it takes a while to get them to the size they can eat them. After 3 or 4 days they are fine but the first couple of days some of the smaller ones will just starve to death without small enough food.
    I hatched about 50 Nothobranchius rachovii MZHL-2005-05 Nicuadala (very very small fry when hatched) and I still give the paramecium to some of the smaller ones. Since they have varied growth rates and I assume it is the males that are growing the fastest.
    Anyway it sounds like you are doing very well with the S. fulminantis I am sure it will make a difference if you can give them small enough food in the first few days.
    I just give maybe a cap full (or about 2 inches of an airline hose) everyday, but I also mixed that up with sifted daphnia and walterworms and green water, whatever is around to keep them happy, then when they are big enough I start giving BBS.

    And here are some pics of my fry raising rack, this shows it in testing - I am getting smarter in my old age and testing such things in the bathroom now. Though the patience is not there yet the soapy look to the water is from plugging leaks with silicon as I spotted them.

    Basically it is a recirculating sytem for fry trays, I got fed up after I lost a few fry the other week when I got caught up with other things and forgot to change water. So this is based on what I saw at an aquaculture department at uni years ago.



    It is just an old pump I had jury rigged to pump water to the top, each tray has a fine filter cut and siliconed to form a partition across the back and behind that air hose glued in about an inch and a half high, so it all just overflows down to a sump/filter at the bottom.



    The interesting thing is it is making lots of oxygenated water which I did not plan on, since as the water overflows it sucks air in and bubbles away. The only annoying thing is it sounds like a mosquito.



    But it so far seems to work well, and was cheap enough to make. I also added extra cloth filters to make sure that fry stay were they should, but with the amount of silicon and stuff I have used I am quite confident in it. I will post more pics of it in action and details if people are interested and when I get more time.


    Scott.
    Thanks again,
    Scott Douglass

  5. #5
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    Quote Originally Posted by Scott_sg
    I will post more pics of it in action and details if people are interested...
    Show me your behind, Scott. No... that should read 'behind the rack', how you distribute and secure the output/overflow tubes.

    Do these tubes punch through the containers or are you using a "double-J" arrangement? For those incoming tubes, I don't see valves to control the flow-rate or are the tubes 'pinched' further up?

    Yours is a mini re-circulating system so be mindful of bacterial outbreaks. Tough to lose a tank of fry, cry when you lose a rack. Since your fry rack is in the bathroom, consider too, a flow-through with slow drip and overflow (discharge to drain).

    BTW, is it just me or did it occur to anyone that the fry containers resemble mini MattenFilter tanks??

    Going back to the culture, how did you manage to sieve the paramecium from the 'microscopic soup' or was your starter ordered online?

    [I'll take a raincheck on the starter cultures as my soup is still filled with goodies but you're free to be further infested with DIY bugs! Maybe that's one reason why folks stop dropping by at my place! ]
    I'm back & keeping 'em fingers wet,
    Ronnie Lee

  6. #6
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    Hi Ronnie,

    You are getting a bit technical for me there I have to admit I am not up on the different filters, but I did pinch some of your ideas from the pics you put up..just on a smaller scale. My main aim was to make sure that fry stay where they are and don't get mixed up or caught in any thing. So a lot of things tended to follow from that.
    My goal was really simple and that was just a drip through system for fry up to about 1cm. So it was as much a prototype and experiment. But it has worked out a bit better than I expected.
    The sump is really a mess at the moment, one section holds the filter media, which is just those clay pipe things and some more filter wool stuff, and the pump is an old dog of a thing I had laying around. It is way too much for there so I just made a bypass (trial and error) to let excess water flow out in the sump for now. But now I know it is working I will invest in a a better pump and sump/filter for the thing.
    I found a 2' glass tank downstairs the other week so I might put that to work for the sump, but that will give a good water volume and be a nice resovoir for the system.
    So I am not going to show you my behind until it is all looking nice
    The pump is just connected to the thicker hose to send it up top and then split into 3 airline hoses to fill the top trays.
    There is probably a lot better ways to do things, but it was really a proof of concept thing. I think I will need to get some one way valves for the top trays though, or rig up a frame to make the output drip into the top, since the only problem as it was set up first was that if the pump was shut off the top trays would back siphon, so at the moment it is chop sticks holding the output hoses clear.
    The trays just lift out for cleaning or if they get too bad easy to replace and make new ones.
    But anyway now I have it up and running I will continue to iron out bugs and improve on things over the next couple of weeks before I try it on any fry.
    I figure the more volume I can put in the sump the better for the water quality and the fry. Since I thought the same as you , I don't want to lose a rack of fry..
    I am not planning on keeping it in the bathroom, but for now it is a good place to test it out in case something goes wrong A long history of &^%^ ups and water all over the place with people yelling at me has taught me something at least.
    I will update more pics when it is all working well but for now I am actually surprised at how well it seems to work, better than I expected!

    My budget roughly was:
    1 shoe rack $17
    2 bags of take away containers $5
    2 bags of the filter media to glue in $5
    4m airline $2
    1 big thing of silicon $5
    1 gravel cleaner $2 - the main lift hose with the suction thing cut off
    1 old pump - but they are cheaper than ever now.

    So it was really just an experiment that so far I am pleased with.
    Scott.
    Thanks again,
    Scott Douglass

  7. #7
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    Hi Scott, seeing your tray system is pretty interesting since I saw a very similar setup over at Ron's some time ago. Pretty much works on the same principles I guess but its a neat way to do things since fry love a slow constant flow of water. Plus, your system puts little stress on them via slow and subtle changes, if that's what I understand it to be.

    Anyway, here's my system, which has worked efficiently for me in most cases at least.



    I use the upper tanks for keeping newborns and the lower tanks as spawning tanks or raising tank for larger sized fry that are able to handle foods like baby tubifex worms.

    I keep the tanks separate from each other since I do not want to risk any sudden disease outbreaks from infecting other tanks. That is why I don't quite like the idea of a flow-through system. With the current sponge filter system I have in use, the bubbling flow of the water onto the water's surface has been adequate for my purposes in terms of oxygenation and also filtration. Keeps the fry happy for days on end. I only need to top up water lost via evaporation.

    About the paramecium, its interesting that you use oats cooked in hot water. It seems that I could culture them too via the oats that I use for sub-culturing my walterworm culture at home, of which the starter was courtesy of Mr Ronnie here.

    I think I'll be needing a paramecium culture soon since I'm expecting some tiny fry to be hatching out real soon..
    Fish.. Simply Irresistable
    Dawkinsia assimilis, Heros efasciatus - Looking for Pelvicachromis taeniatus
    -back to old school fish-

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    I do not have Paramecia right now, but wish I did. Pure cultures are hard to come by, as most hobbyists are a bit careless and usually contaminate their culture.

    I kept a pure one going for a number of years, in Fremont. I used about a 1L jar that had a large glass lid with polyethylene gasket that made a good seal by just setting the lid on the jar.

    Some folks use dried corn husks, but, as I recall, I just had a bare jar that was fed a drop or two of Liquifry No. 1 every day or two when I was actively using the culture, and about every week when it wasn't needed.

    I had a baby medicine dropper in an unusual color that was the only thing in my fishroom to ever touch the culture water. Top-up water, to replace that used for feeding, was from a carbon-filtered storage barrel that was well covered and kept very clean. Since this was the prime potential for contamination, it was used very infrequently. I was careful to be sure no tank water containing other larger-than-bacteria organisms ever contacted the culture.

    That original strain was obtained from a fellow BAKA member, who probably got his starter from Carolina Biologicals. It lasted for years without anything growing in it but the original strain of 2-300 micron Paramecia. They were big enough to see as individuals in a bright back light, and the jar was definitely cloudy when being fed enough to be useful as food.

    I usually put a dropper of culture on a wad of Java Moss in a baby container and then one or two drops of Liquifry No. 1. In a day, a cloud of critters could be seen near the moss, and enough were scattered around to be in the face of new babies needing to develop that snapping reflex. The filter-feeding Paramecia kept the water ultra clear, except for the cloud they formed with their own bodies.

    Bottom line: Paramecia are easy to keep and use, but watchful care is needed to avoid contaminating them from a critter source like tank water. Rotifers and other animals will quickly out compete them and you will not have a good culture if you are not very careful.

    Wright

    PS. If your culture gets contaminated, as quickly as possible start a new one by selecting a few individuals under a microscope with a good pipette, and transferring them to a new jug. One drop of Liquifry and you'll have a new culture in a week or less.
    01 760 872-3995
    805 Valley West Circle
    Bishop, CA 93514 USA

  9. #9
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    Hi to everyone, sorry for the late reply, I decided to take myself out on the town for the weekend, which I have not done for a very very long time. Problem is it is getting harder and harder to recover each year..

    The Paramecium are actually very easy to culture, but like Wright says it is easy to contaminate them. In the home you will never get perfectly sterile conditions, so the idea is to use some common sense and try and make it as favourable for only one type of organism to grow. This means I always keep a few cultures going.

    For example, I never lift the lid completely off a culture, rather just lift it partially up to slide whatever I am using in, get what I want and then close it again. In short the more exposure the culture has to the air or dirty equipment the bigger the problems will be. For feeding I simply use a short piece of tubing and dip it in about an inch or two then hold my finger over the end. But I do rinse it in boiling water before I use it and then work fast to take what I need, and put it in a dish

    It is much better to take however much you need in one go rather than dipping in and out all the time. It is not really that much of a hassle really just a matter of being sensible. But in the home it is only a matter of time before you infect a culture so that is why you should always have a few going.

    The great thing about the Paramecium is once it is up and running, the less that you touch it the better it will do. The Paramecium themselves feed on bacteria, so the cultures are almost self sustaining. The traditional culture medium for them is wheat, but since I use oatmeal for my walterworms I simply use that for my paramecium. But really any cereal will do. Occasionally to give them a boost I will give them some green-ish water. This is the water from the pot when I have boiled vegetables, I simple pour it hot into a clean container and seal it.

    The oats themselves are not cooked, rather i always add the oats to a clean container filled with freshly boiled water. This sterilises (within reason) the setup so that once it is cool I can then innoculate it with some culture.

    For Ronnies question which I missed the other day as to how to seperate them out. There are really lots of ways to do it, the classic lab way is a glass tube with some wool in it. The paramecium are one of the fastest of the common protists so they will swim through it the fastest. Picture a maze for them I guess. I simply made a partitioned container with filter wool in the middle added green water to one side and then a few hours later took clear water from the other side. I then diluted that again and innoculated a dozen or so sterile cultures I had made. Eventually with a bit of luck and playing around I was able to end up with a reasonably pure culture to work with. So then once I got that I then basically followed the above steps again.

    The biggest danger to the Paramecium is actually other Protists, Didineum sp. are very bad, (Dont' quote me on the spelling) since they feed exclusively on Paramecium and will quickly eat all the Paramecium. But using the above methods it is tedious but not impossible to seperate them all out. For me I get a kick out of it so I dont mind seeing what I can purify and play around with. But in general it is a matter of reserching what you want and then thinking of a way. For example Euglena sp. can be seperated out by simple using a light, but then you will get other photosynthetic animals, so then you need to refine it and so on. The standard microbiology way of doing things is to dilute things down so that you are down to one "animal" per drop say and then use that to innoculate cultures.

    So there is really no great big mystery to it, rather just some patience and interest in the creatures. Once you have a pure culture they are easy.

    My fry rack I hope to continue work on this week, but my own preference for fry has been smaller containers, I like the more control I can have over them. Since there are plenty of times I have had a lot of fry just dissapear in a larger tank. But that is just me. I understand the disease risks of a circulating system for raising fry, but I think any method of raising fry will have advantages and disadvantages. In the end if my idea turns out dumb, it was quite cheap to set up and at the worst I could see it would be good for keeping my live food "alive" in between my trips to the local fish shops.

    Scott.
    Thanks again,
    Scott Douglass

  10. #10
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    Quote Originally Posted by Scott_sg
    There are really lots of ways to do it, the classic lab way is a glass tube with some wool in it. The paramecium are one of the fastest of the common protists so they will swim through it the fastest. Picture a maze for them I guess. I simply made a partitioned container with filter wool in the middle added green water to one side and then a few hours later took clear water from the other side. I then diluted that again and innoculated a dozen or so sterile cultures I had made.
    Hi Scott,
    thanks for this and your other most informative and interesting posts on culturing "wee critters" !
    I will have a go at that technique when things warm up in the spring next year.
    Meantime,, you say "there are really lots of ways to do it," I wonder if you have any other interesting (web?) refs ?

    I have a microscope and have observed (in an amateurish way!) paramecia, volvex, animacules, rotifers, tardigrades - facinating!, etc
    but I lack any education (and equipment techniques) in micromanipulation of these things

    Edit later :
    Ooops, sorry folks, I forgot my sig. :
    Malcolm,
    in cold and wet SW England.

  11. #11
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    Hi Malcom,
    Glad to know it was of some use to you, sorry for the late reply but I seem to get caught up in a thousand and one other things.

    Separating them out, is probably not as easy as I have made it sound, in the home it takes a lot more trial and error. But I don't mind as I enjoy seeing just how far I can go with ordinary gear around the house. And the creatures are interesting in their own right.

    For web links, there is not much I have been able to find on the actual handling of the organisms, although there are some good pictures online. Even the text books tend to gloss over the actual techniques, since most of it is really taught in the lab. Although if you ever see any of the older text books, grab them, for some reason a lot of the older books seem to explain more hands on stuff -50s 60s, even though the names will change (and they still do today anyway) they are great if you come across them in second hand shops.

    For Separating them out it really depends on what you want to separate and purify. If you are just playing around for fun, then an easy experiment is simply to buy a dozen or so containers and add different ingredients to the water in each, one yeast, one wheat, one grass, one bicarb - whatever you have around. Some can be kept in light others in dark and so on. After a week or two you should see different creatures will favour a different environment and essentially take over that container. So there are lots of things you can do with just some pond water.

    *** WARNING IF YOU HURT YOURSELF DON'T BLAME ME - AND NO POINT SUING ME, THE EX WIFE ALREADY TOOK ME TO THE CLEANERS ***

    If your microscope is ok, then you can always make micropipettes by begging borrowing or stealing some thin glass lab tubing. You can always buy the stuff but probably easier just to ask around. Anyway heat it in the middle rotating gently in a little spirit burner (can make your own easy also) and when it starts to soften pull the two ends apart, they will pull to a very fine thread before breaking, then simply break off at the length you need.

    With some practice you can make very good pipettes to then individually manipulate individual animals under the microscope and use them to inoculate other cultures etc.

    I enjoy playing around with these things, so it is a little sideline for my killifish - and occasionally gives good fry food. But don't be afraid to play around and experiment. All that is needed is some commonsense and some interest. Unfortunately schools seem to knock both out of kids.

    And the SW of England is not so bad, unless they have you running up and down the Dartmoor at night, look on the bright side you could be in the NE of England and it is colder than Scotland there I have a bad memory of sleeping outside the train station in Newcastle one night, but that is another story.

    Scott
    Thanks again,
    Scott Douglass

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