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plante1999
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Thanks for the tip!, As you can see I'm not a microbiologist.
So. my 500 ml solution is clear but have some carbon dust in it. But no cloudiness like previous try. I guess the bacteria are in the activated
charcoal. I added 1 ml 20% ammonium chloride solution as the ammonium source, and the pH is still 8, which is supposed to be ideal for them. I have
about 2 g of sodium bicarbonate buffer. When the ammonium will be used up, HCl will react with the bicarbonate, making chloride and the nitric acid
will too. When the bicarbonate will be used up increase in acidity should be observed and prove biological activity.
Soon I will try to use urea for the ammonium salt source and add calcium carbonate buffer to the bottom. I may change the substrate too.
Phlogiston if other organism try to live in there it should still be good since these bacteria usually live in these condition ( fish tank).
[Edited on 17-12-2012 by plante1999]
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phlogiston
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There may be other bacteria that do well under the same conditions that do not produce nitrates, and they may take over your fishtank.
Where did you get the composition of the medium? You don't seem to add any source of carbon. Most bacterial media that I have ever used contain
glycerol, glucose, ethanol or some other 'fuel' that bacteria like.
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plante1999
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Nitrobacter do not use carbon source, they will only grow with CO2. In the presence of organic compound they will not grow. And by the way I don't use
a fissh tank but I mean these bacteria could be found in it.
Edit:
The pH dropped to 7, so I suppose there some activity in the medium, more update soon.
I bought calcium carbonate and urea, I will do an urea addition.
Edit 2:
I added 1 ml of saturated urea solution and the pH turned back to 8. I may add this peppermint calcium carbonate pills soon.
[Edited on 17-12-2012 by plante1999]
[Edited on 17-12-2012 by plante1999]
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plante1999
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Few tenth of minutes after the urea addition the pH was 7.5. It was run for another hour and then the pH was found to be 6-7 so I tested my calcium
carbonate pills, I dropped one in distilled water to see if there was an evident reaction. Nothing happened. Then I added 9 pills of 0.5g calcium
carbonate each, or 4.5g. Imediatly bubble formed on them, they dissolved somewhat and 2 minute latter the pH was back to 8-8.5.
[Edited on 17-12-2012 by plante1999]
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Mailinmypocket
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Peppermint? Are you using antacid tablets?
Why not use chalk?
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plante1999
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Yes I use antacid tablet because I don't have any chalk and I didn't wanted to use seashells.
Edit:
I added one ml of saturated urea sol., the pH is still 8 and the calcium carbonate is partially dissolved. I should be able to test for calcium salt
by taking some of the solution and adding sodium bicarbonate solution to it. Cloudiness would tell that some soluble calcium salt are present.
Summary of the first 24 hour of run of the bioreactor:
hour 1: Ammonium chloride, sodium bicarbonate, activated charcoal and the bacteria source were added. pH 8
Hour 11: The pH was now 7. First urea addition, 1 ml of 26%. pH 8.
Hour 16: pH 6.5-7. Addition of 1ml of 26% urea and calcium carbonate. Effervescence with the calcium carbonate. pH increase to 8.
Hour 20: pH still 8. Addition of 1 ml urea 26% sol. Calcium carbonate mostly in tiny pieces format.
Hour 22: pH still 8. Addition of 1 ml 26% urea sol.
Interpretation:
Hour 11: The bacteria colonies used most of the ammonium to make nitric acid and most of the bicarbonate was neutralized. When the urea was added,
urea nitrate was formed neutralising the acid and increasing pH.
Hour 16: Urea was metabolised by the colonies to make nitric acid lowering the pH. Calcium carbonate reacted with the nitric acid to yield CO2 and
Calcium nitrate, increasing the pH. The calcium carbonate buff the pH to 8.
Remark:
I read that the calcium carbonate is not only used to buff the pH, but also as the carbon source for the bacteria, as air CO2 is usually insufficient.
I wonder if the pepermint, dextrin, starch and sucralose interfere with the bacteria.
At hour 33 I will test for the calcium content of the solution.
Edit:
I tested the calcium content of the solution with sodium carbonate solution. The solution was very slightly cloudy. I don't know if it is the original
solution or the reaction that made it cloudy. Result: Inconclusive.
But even if all the urea had bean turned to nitrate, only 0.1% of the solution would be calcium nitrate.
I tested the filtered solution with the oxalate test, it was more precise and some calcium is present in the solution as very slight cloudiness
appeared. To get a clearer test the bioreactor must run way longer
[Edited on 18-12-2012 by plante1999]
[Edited on 18-12-2012 by plante1999]
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plante1999
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Ok, so I have designed two setup. I would want to know which one would be the best. The first one is the currently running one. One is in water
medium, and the other would be moist only. both get air from an air stone.
1:
Green is the air stone and the pale blue the tube. The white is calcium carbonate and the black is activated charcoal. Water medium.
2: It is a variant of old nitre bed. The orange is a clay plate and the white potassium carbonate. The purple is calcium carbonate pieces and/or
activated charcoal. The set up is only slightly moist and air come from the air pump.
Thanks!
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plante1999
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Ok, so I found that the antacid contained more than 2 time the weight of calcium carbonate in sugar, starch and other thing like that. To be sure my
results were not influenced by the presence of these undesirable compound I made a new colonie in a new medium type.
The plate is made from tera- cotta, onto it there is a calcined calcium carbonate layer onto which the pump is placed. On the pump there is broken
seashells. I vaporize dilute urea solution onto the seashell to keep them wet and I added bacteria seed. I hope that calcium nitrate creep out of the
plate as crystals.
<iframe sandbox width="560" height="315" src="http://www.youtube.com/embed/jfarUfBYIMQ" frameborder="0" allowfullscreen></iframe>
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plante1999
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I don't want to spam SM more with my posts. So, if someone is interested by this experimentation, go on the nitrate from ammonia tab of my website. If
you have any questions you can ask them on my website. If you want to try please write your experimentation in this thread.
Have a good day!
http://hclo3chem.weebly.com/
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watson.fawkes
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Experimental progress isn't spam. That remains true even if
folks aren't replying with each update. I've been reading, though I have little to say. (Though since I rarely watch video, video updates are mostly
the same as no updates for me.)
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plante1999
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Quote: Originally posted by watson.fawkes  | | Experimental progress isn't spam. That remains true even if
folks aren't replying with each update. I've been reading, though I have little to say. (Though since I rarely watch video, video updates are mostly
the same as no updates for me.) |
Thanks for the comment, however I have the feeling I'm spamming SM. I take good note for video. I did a videa because I cannot do good pictures the
only camera I have is a webcam, a picture with a webcam is not very good. So you would need to try to imagine whatever I say.
By the way you can still read what I do on my website but here a summary of it:
My new design doesn't have any soluble calcium, but does have a lot of urea. I will wait until the 31 to see if any good result come out of this
design. I still think that urea SHOULD be metabolized as most protein breakdown produce urea if a remember, and these bacteria feed on protein waste.
Still I will try to find a good ammonium salt for them, without a deranging anion. I will also try to find a more porous media.
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watson.fawkes
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| Quote: Originally posted by plante1999 | | I did a videa because I cannot do good pictures the only camera I have is a webcam, a picture with a webcam is not very good. So you would need to try
to imagine whatever I say. |
You've involved yourself in communicating chemistry. You'll succeed well only if
you can both do chemistry well and communicate well. Part of modern communication is multimedia production. Therefore to further your efforts, balance
your budget between chemistry equipment and communications equipment. (In particular, you can go a long way with good lighting and a not-so-great
camera.)
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plante1999
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I bet in the 18th century there was no need for pictures... I will try to find something, but I get something like 100 CAD, just enough to restock my
lab, If I get a camera, important reagents would be missing of my lab. Argggg. So anyway, does anyone have an idea of an ammonium salt, which
solutions are of pH about 7 and that the anion would not interfere?
[Edited on 20-12-2012 by plante1999]
[Edited on 20-12-2012 by plante1999]
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Vargouille
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Ammonium acetate might be your best bet. Ammonia and acetic acid have the same value for Kb and Ka, respectively, so the pH
should be about 7.
EDIT: I'm not sure if acetate will interfere, mind you, so that'll take some testing on your end.
[Edited on 20-12-2012 by Vargouille]
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plante1999
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Thanks for the idea! However I'm not sure if acetate would kill the bacteria, as they don't support organic material. If I had ammonium nitrate, it
would be perfect. I'm wondering why they don't metabolize fast urea, in nature proteins decompose to urea mostly if I remember.
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phlogiston
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Ammonium sulphate and Ammonium chloride are common in bacterial and yeast media.
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"If a rocket goes up, who cares where it comes down, that's not my concern said Wernher von Braun" - Tom Lehrer |
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Vargouille
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There is some evidence that a Nitrobacter specimen would be fine with acetate. It uses it as a carbon source, apparently. There is also a mention in this article that acetate does not harm Nitrosomonas bacteria, and is also used as a carbon source. This is fortunate because not only is ammonium
acetate a (likely) neutral salt of ammonium, but it would appear that it is also compatible with the bacteria.
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plante1999
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Sound rather good, I just don't want a 30% bi weight acetate in the nitrate produced, but if they metabolize it, it should slowly be used up. At least
they are compatible with the salt.
Alright, then I will test using ammonium acetate. First I will need to make some ammonium acetate tough.
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plante1999
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Urea is slow to metabolize by the bacteria. This may be explained by the very low dissociation into ammonium and cyanate. Even the ammonium must
dissociate into ammonia and H+ because the bacteria use only free ammonia. Such low ratio of free ammonia slow the process a lot. Free ammonium salt
have a very good dissociation constant, which give very fast conversion to nitrite. However urea might have a very good advantage, it have the ability
to make hydrogen bond. As such it can act as a very effective buffer for the medium by making urea nitrate. After some time, this urea nitrate would
be metabolized all the way to nitric acid and CO2. More urea would probably bind the nitric acid produced too.
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plante1999
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When I tested this morning, the pH was 6-7, I do not explain this since the seashells where supposed to neutralize the acid. So I added fee drops of
sodium bicarbonate sol. to brink the pH to 8 and then I tried to add one ml of saturated ammonium sulphate. After a few hours the pH was 7 and there
was a very slight ammonia smell. This definitively prove that ammonium salts are metabolized WAY faster than urea. Soon I will make ammonium acetate
and use it as the ammonium source.
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plante1999
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From my results of my new design I think I misunderstood what happened in my first design, I made a revision of it, here is it:
Summary of the first 24 hour of run of the bioreactor:
hour 1: Ammonium chloride, sodium bicarbonate, activated charcoal and the bacteria source were added. pH 8
Hour 11: The pH was now 7. First urea addition, 1 ml of 26%. pH 8.
Hour 16: pH 6.5-7. Addition of 1ml of 26% urea and calcium carbonate. Effervescence with the calcium carbonate. pH increase to 8.
Hour 20: pH still 8. Addition of 1 ml urea 26% sol. Calcium carbonate mostly in tiny pieces format.
Hour 22: pH still 8. Addition of 1 ml 26% urea sol.
Interpretation:
Hour 11: The bacteria colonies used some of the ammonium to make nitric acid and most of the bicarbonate was neutralized. When the urea was added,
urea nitrate was formed neutralising the acid and increasing pH.
Hour 16: The remaining ammonium was metabolised by the colonies making nitric acid lowering the pH. Calcium carbonate reacted with the nitric acid to
yield CO2 and Calcium nitrate, increasing the pH. The calcium carbonate buff the pH to 8. The urea probably did not reacted, or very slightly.
In my last desing I added ammonium sulphate, and I smelled ammonia after a tenth of minutes. I suppose this reaction happened:
(NH4)2SO4 + CaCO3 -) CaSO4 + NH4HCO3 + NH3
I will try ammonium bicarbonate and ammonium actetate. I know a preparation to make the ammonium bicarbonate, but does anyone have another, or have
already tried?
Making the odor. I may do two medium at the same time. This product explixitly say that it contain nitrobacter and nitrosonomas:
http://www.fritzzyme.com/index.php?p=fritzzyme-nitrifying-ba...
They also sell ammonium chloride to use when preparing the aquarium with the fritz zyme 7. This prove the content of the bottle. They seams to know
what they make, I would probably prefer this product then the walmart one. They also have the concentrated product too which is more interesting for
me I will probably buy this too, or something similar:
http://www.ebay.ca/itm/Aquarium-Fish-Tank-Super-Sponge-Bio-F...
And I will probably need something to test nitrates, like this:
http://www.ebay.ca/itm/AQUARIUM-PHARMACEUTICALS-TEST-KIT-NIT...
Does anyone know how I could glue acrylic to terracotta? I will use acrylic to make a square tube and then glue it on a terracotta plate. I will put
the bio filter in the square tube and put water. With this set up, it will be easier to make experimentation. I hope, like the second set-up, that
crystals of nitrate will creep out of the plate.
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watson.fawkes
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Epoxy should work. Acrylic is very friendly to
chemical adhesives, and terracotta is porous enough that any gap-filling glue should work. Just make sure you have adequate contact area. Add some
acrylic flange for extra area of the cross section of the walls of your tube aren't enough.
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