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Amoeba Culturing

About microscopic forms of life, including Bacteria, Archea, protozoans, algae and fungi. Topics relating to viruses, viroids and prions also belong here.

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Amoeba Culturing

Postby amoebaslayer » Tue Jul 24, 2012 4:24 am

Wanted to join this forum to ask a question because I haven't seen much online about culturing for amoebas. What I do on occasion is boil water (mostly pond water) and then add beef bouillon cubes or cooked rice. Sit this outside for a few days and then take a sample of the water. Most of the time I get amoeba's without a problem doing this. Here's my concern and where I need some help. When looking at these samples the other day I noticed something a little different about the amoeba. So I jumped online and started reading. Now keep in mind I've been looking at these samples for months. So when I was reading online 3 days ago I ran across something I had no idea existed "naegleria fowleri" or "brain eating amoeba" :shock: . I keep reading the information about it and I have come to the conclusion that this amoeba is everywhere. Thing is the disease it causes is really rare (PAM). I also read where 2 kids last month died from this while swimming in lakes. Basically, your fine unless you somehow sniff this stuff up your nose and I'm not convinced yet as to how far this stuff has to get into your nose before it turns bad things worse. I read somewhere that once it is in your nose it senses the nerves communication and swims towards the nerves that make you smell things. Anyways, so the strange movement or the different amoeba that I'm seeing could possible be Naegleria f. but I'm not sure. I can't find anything online about it being possible to culture Naegleria f. Does anyone have information about this amoeba other than CDC report? Is it possible to culture this by accident or maybe not by accident but when culturing for amoeba p. the Naegleria show up? :?:
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Postby david23 » Wed Jul 25, 2012 12:38 am

the issue here is that even if you are able to the culture this in your sample, how do you know this is the amoeba you want? You still have to do so many PCR, sequence for the particular genes etc etc. After all they would just look like normal amoeba under the scope.
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Re: Amoeba Culturing

Postby amoebaslayer » Thu Jul 26, 2012 4:25 am

well.. I though about that. What I've found (but haven't tried yet) that when you introduce distilled water the N.F. amoeba transform and grow tails (Flagellate) so that they can swim away.
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Re: Amoeba Culturing

Postby david23 » Fri Jul 27, 2012 1:37 am

what kind of lab do you have, I can see how sequence DNA might be a little costly. So how about this, do you have lab mice to test? Does your lab have permission to do this.
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Re: Amoeba Culturing

Postby amoebaslayer » Thu Aug 02, 2012 1:58 pm

no lab.. just doing this at home. I had microscope already and never really used it and one day I decided to look at my tap water. I took biology in school but I didn't really care about it much. However, the more I read about this the more I would rather not try to make it appear. I would like to see the amoeba just to watch them move and feed. Recently, I read the "Cultivation of Pathogenic and Opportunistic Free-Living Amebas" and that pretty much did it for me right there. I also wonder what else is out there we don't know about as far as what may or may not be "pathogenic" to animals/humans. I do have one question about the word "culturing". When you create a culture of some kind, does this mean you already have a sample of something and you are trying to grow it or does it mean you are creating a culture so that something appears in the culture medium?
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Postby Atropa » Fri Aug 03, 2012 4:39 am

Where are you doing your culturing? N. fowleri is usually found in stagnant, warmer waters, in warmer climates. It's probably not something you want to play with if you have it... There are a lot of photos and descriptions you can probably use to figure out if that's what you've got swimming around in your sample. N. fowleri is capable of a flagellate state, not seen in most other amoebae, and it can apparently be induced...
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC221533/
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Re: Amoeba Culturing

Postby amoebaslayer » Fri Aug 03, 2012 3:07 pm

2 places I've seen this amoeba (not sure if it is naegleria or not). I have an old barrel that is metal that stays full of water most of the time. It has been sitting in my yard for about 2 years now. At one time someone used the barrel to burn trash. The other place was a near by very small pond that cows drink water from. I felt the urge to go check out that water after reading about amoeba's. I live in Texas very close to the coast near Louisiana. Here is a video of what I am seeing and I would like to hear some input on what it may or may not be. I have also boiled up some beef broth and let it stand outside for 2 days and some of the same thing has shown up.

http://www.youtube.com/watch?v=RX-dUUKG ... e=youtu.be
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Postby Darby » Tue Aug 07, 2012 1:52 am

I worked for years in one of the few labs that dealth with this. It's a very common ameba, but extremely rare pathogen - even with access to the olfactory epithelium. Humans are not a good ecosystem for them - we used to culture pathogens in mice, which are very susceptible.
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Postby amoebaslayer » Wed Aug 08, 2012 11:33 pm

Thanks Darby - Wish there were more information on this amoeba. I've tried to read a lot of material online and my thoughts are that this amoeba doesn't make it up the noses of many people making it a "rare" pathogen. Not really how sure it is in our environment because I've only see that amoeba in the video in 2 places. An old barrel and a very small pond.
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Postby Darby » Sat Aug 11, 2012 11:47 pm

The assumption is that many people get exposed, but the ameobas are rarely able to penetrate the surface and/or survive in the tissues. When mice suck them up, they always get infected. One of the folks in the lab was trying to figure out what the Naegleria - human incompatability was, but I don't believe he got close.
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Re: Amoeba Culturing

Postby amoebaslayer » Mon Aug 13, 2012 8:48 pm

Yeah, I think that is the question. Mice being induced with this get infected and from what I read so do sheep and other animals. I don't think there will be any human volunteers to study for this type infection. How are the mice induced? Were they spray mists or more of a forceful flow in the nose?
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