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Old 01-03-2014, 06:34 PM   #231
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Going back to the paper about Nitrospira from Dr. Tim. I like this paper because it has a nice graph about the entire nitrogen cycle length, and also made a point to detect when certain bacterias appeared. This appears to be one of the few papers out there that basically just studies "cycling" (although its primary purpose was to prove Nitrospira, its secondary effect was a study on the cycle)
Nitrospira-Like Bacteria Associated with Nitrite Oxidation in Freshwater Aquaria

"Time of Nitrospira-like bacterial appearance.
The daily concentrations of ammonia, nitrite, and nitrate over the first 33 days after setup of a new aquarium are presented in Fig. ​Fig.6.6. The trends were as expected, with ammonia peaking about day 12. Nitrite values increased starting at day 12, peaked at day 21, and decreased to below detection limits by day 26. Nitrate values steadily increased from about day 15 onwards. DGGE showed that the band corresponding to clone 710-9, the putative NOB, first appeared on day 12, with the relative intensity of the 710-9 band increasing daily based on relative fluorescence units of rDNA amplicons"

One important thing to note, as you look at the graphs, is that all of the additions of ammonia are in mM. So that means when it says 4mM ammonia it means 68ppm ammonia. NitrIte in this study reached 11mM, or 506ppm, before dropping.
Again: lab work is so far outside of our scope and does not stall. Our cycles will not stall due to levels of ammonia or nitrite that we encounter. They may stall due to OTHER THINGS but not for that SPECIFIC reason.

"Time series.
Three aquaria were set up as previously described with 4.53 kg of gravel and were filled with 30 liters of city water which had been passed through activated carbon. The test was run for 138 days, during which the aquaria were individually dosed with 8.9 mmol of filter-sterilized (0.2 μm) ammonia (as ammonium chloride) on the first and second days of the test. From days 12 to 78 of the test, further additions of 8.9 mmol of ammonia were done on average every 3 days. A total of 246 mmol of ammonia was added to each tank during the test. The water was sampled three times a week for chemical analysis. The aquaria were run for 80 days with freshwater, at which time the water was switched to seawater (32 ppt) by draining and refilling with water mixed with artificial sea salts (Marineland Commercial Aquariums, Moorpark, Calif.). After the switch, the testing continued for an additional 57 days."
Total of 246 mmol ammonia added. 4189ppm ammonia. lol!

"Time of NOB appearance.

Three all-glass aquaria were established as described above. A 34-liter sample of city tap water, which was passed through activated carbon, was added to each aquarium, which contained 4.53 kg of gravel. Initially, 0.71 mmol of filter-sterilized (0.2-μm-pore-size filter) ammonium chloride was added to each tank, followed by an additional dosing of 5.0 mmol of NH4Cl on the fourth day. On days 10, 15, 18, 23, and 30, further ammonia additions of 8.9 mmol were made to each aquarium. During the test, a total of 50.4 mmol of ammonia was added to each aquarium. Water samples were collected daily.
Two 10-g samples of gravel were collected from each aquarium daily for 33 days."
This one had an initial dosing of 12ppm ammonia with 4th day dose of 85ppm and maintenance doses of 151.6ppm.
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Old 01-03-2014, 07:31 PM   #232
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So mM MEANs moles per liter.
So mMol or mM conversion to ppm is exceptionally simply. In practice it is nothing more than the concentration of the solution * the molecular weight of the solution.
So if they say "50 mM of Ammonia" you just multiply 50 * 17.031 (molecular weight of ammonia) to get 851.55 mg/liter which is 851.55ppm

Source: http://www.ehow.com/how_8412601_conv...moles-ppm.html

ppm = A x mmol/l
mmol/l = ppm/A
Where A = atomic mass of the ion

mmol/L is millimoles per litre
mM is "milliMolarity" which is millimoles per litre
both are millimoles of solute per litre of solution
So we may see it written either way but it means the same thing.
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Old 01-03-2014, 07:39 PM   #233
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Originally Posted by threnjen View Post
LOL Caliban! In one post you're trying to keep us on track, in the next one you're off on the irresistible tangent

This is all so interesting, it's hard to keep on track!

I agree with your post a few back - we need to come up with the test parameters for the volunteers. My husband advised me that first I need to make a chart of all of the "known" parameters relating to bacteria growth, using all of our sources. i.e. temperature (not just the ideal temps - how slowed is growth outside of them?), rate of doubling of colonies, proportion of colonies in fully cycled samples (I HAVE seen bits of this, somewhere, about the proportions) etc, ideal DO, etc. Then from there we form our hypotheses. He said that I have seen enough data that I should be able to easily run a simulation of what I would expect to see given different dosing scenarios. I will try to work on this today and post them.

As far as hypotheses, I *think*, and I can only say I think as I need to think it through more, that ultimately *my* hypothesis (I believe yours will be different) will be that:
a) The level of dosing does not influence the cycle length, but does influence the end nitrification capacity
b) Water changes can be eliminated through manual additions of phosphorus (fish food) and baking soda at cycle start
c) Cycles without water changes will complete faster than cycles with water changes
d) I feel uncertain on this one. But I'll say it. After initial dose is reached, no additional ammonia is required, and will only prolong the cycle.
e) No levels of ammonia or nitrite in our scale of application will stall the cycle

To address these in turn:
a) I'm not on board anymore that EITHER your .25 dose will end faster OR that my madcap 18ppm dose will end faster. I think it all ends up the same because I've seen so many lab graphs charting the cycle. To that end I think it will take exactly the same-ish amount of time to complete the cycle for these dosings and therefore a larger dose will prove to be more efficient
b) We only water change to stabilize pH and phosphorus, but these readily available additives make that irrelevant, and affect c
c) I think that the cycle is slowed by water changes because you are removing substrate that the nitrite oxidizing bacteria need to eat and multiply. If all is left alone, I propose the ammonia eating bacteria will produce exactly the correct amount of nitrite that the nitrite eating bacteria must consume in order to create a perfectly balanced population.
d) Why do we dose back up? Is it because we fear the ammonia eating bacteria die? Or is it because we want to be sure that the bacteria can successfully eat Xppm all at once (rather than the same individual bacteria eating some every day). If it's the latter, then a higher initial dose will do the job. aka Dose it to 8ppm and never add ammonia again. If it's the former - they don't. We have proven this.
e) I simply believe this to be true after all of our work
Ok i agree with some of this.
a) i have to admit im leaning more towards the conclusion that it takes x amount of time to colonise bacteria so deal with it.
b) no need to change water agreed.
c) Agreed
d) i dont really know. Fear? fear that bacteria will die out one dosing of 4ppm would be enough IMO. remember bacteria was still able to consume lab levels of ammonia months later.

We can however not assume that low ammonia dosing would be faster or slower. It hasnt been donr this way in labs?

Do we even have to bother with conditions? afterall these lab tests would have been carried out in optimum conditions anyway. I can say that your goal has been fulfilled. You do not need to change water during a cycle. Patience is key.

we have established useful things that help. Maybe we could suggest that fishless cycles require a water temp of 30 degrees with a close to neutral pH in a dark room. one test with 4ppm ammonia and the next starting at .25
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Old 01-03-2014, 07:42 PM   #234
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Can you try it with a case already worked out? I just had a quick look to what it had and it looked a nice find.
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Old 01-03-2014, 07:54 PM   #235
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Quote:
Originally Posted by threnjen View Post
So mM MEANs moles per liter.
So mMol or mM conversion to ppm is exceptionally simply. In practice it is nothing more than the concentration of the solution * the molecular weight of the solution.
So if they say "50 mM of Ammonia" you just multiply 50 * 17.031 (molecular weight of ammonia) to get 851.55 mg/liter which is 851.55ppm

Source: How to Convert Millimoles to PPM | eHow

ppm = A x mmol/l
mmol/l = ppm/A
Where A = atomic mass of the ion

mmol/L is millimoles per litre
mM is "milliMolarity" which is millimoles per litre
both are millimoles of solute per litre of solution
So we may see it written either way but it means the same thing.
yes i see this math. so 851.55ppm of ammonia would = to about 2299.185 nitrite! lol

so one dosing of 4ppm ammonia would equal a total of 10.8 nitites throughout the whole of he cycle? since we read that even a 90% water change only removes 30% nitrite this could be a reason we nitrite readings remain 'off the charts' after 4 50% water changes that are often encouraged. even after that nitrite would be dark purple.
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Old 01-03-2014, 08:13 PM   #236
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Originally Posted by Delapool View Post
Can you try it with a case already worked out? I just had a quick look to what it had and it looked a nice find.
I don't even think we could do it on such a large scale. I am just not sure how we could even measure the PPM required to match this. The API test isn't made for such a large test!
I think at *LEAST* we can have one of the test parameters be what anyone in aquaria would consider an "outrageous" dose of ammonia, like 20ppm, which can't even be measured directly but can be added by finding the correct amount for 1ppm and doing 20x

OH! Caliban - The tests from the Dr Tim paper also used aquariums, even with these OUTRAGEOUS doses, implying to me that there is PLENTY OF PHYSICAL SPACE to colonize. Agree?

Also - I bet it can be mathematically estimated the amount of ammonia to add at the beginning if you want your end nitrification capacity to be x. Although I have failed to find the EXACT rate of conversion of ammonia->nitrite->nitrate expressed in time, we might just have to assume it takes the same amount of "time" for a single bacteria to eat and excrete, and from there just use the doubling time of the bacteria to determine this. So like if nitrosomonas doubles on avg every 10 hours and nitrospira every 22, can we use this information to estimate the amount of initial dosing of ammonia that will ultimately grow NOB capable of converting 4ppm ammonia in 24 hours.
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Old 01-03-2014, 10:42 PM   #237
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I found this interesting thread on another forum from 2008 where this person "proves" that waterchanges improve the cycle.
However I hypothesize that what the water changes actually did was replenish phosphorus, dissolved oxygen and pH just enough to help the cycle complete ONE DAY faster.
Overall I find his conclusions statistically insignificant. To get a true matching conclusion, he would have needed to consider these other variables and "dose" them to his no-water-change tank.

Why Water Changes During Cycling Are Good - Tropical Chit Chat - Tropical Fish Forums

Edit: ultimately I guess he was just graphing some function he wrote?
How does that function take all the other things that the bacteria "eat" into account? I guess it just doesn't make sense to me.
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Old 01-03-2014, 11:34 PM   #238
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How is it that these conversations always break out when I'm on vacation out in the woods somewhere without internet? There's so much stuff that I would have liked to comment on, but now we're 200 odd posts in, it's hard to really comment on anything specific.


So right now, exactly what are you trying to answer? That is to say, what it your current line of inquiry?
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Old 01-03-2014, 11:38 PM   #239
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WHY HELLO. Your forum handle and cute picture of Bill Nye tells me you will be a delightful addition to our thread!

I just tried to consolidate our purpose to my father-in-law, so let me post what I told him and perhaps that will tell you our "question"
"Even though the process of artificial colony building (called "fishless cycling") is quite popular, there are heavily varied recommendations for how to correctly perform this task, and no standard practices. There are also a lot of perpetuated myths/inaccuracies (which I can see without even being a chemist/biologist), and even worse people "stall" all the time in their cycling attempts which leads to a lot of frustration. So on a forum, some other other people and I have been poring over scientific journals in an effort to better understand the cycling process (the name for the establishment of this complete bacterial colony) and eventually have enough scientifically validated data to present our closest estimate of a "standard practice" for fishless cycling which will yield success in 99.99% of cases."

What we want is to:
* invalidate perpetuated myths about fishless cycling
* produce cycling instructions that will actually work in most cases
* produce the fastest possible cycling instructions
* as a side-project, improve information about fish-in cycling

Some of the major hypotheses just one page back on page 23.

If you really have a good grasp for science, I'm sure you could seriously help us. I think the heavy stuff starts around page 12?

Also, we're not claiming we're discovering a bunch of new things, but we ARE finding actual scientific studies and citations to back up some of things we read about cycling.

Also please do comment if you see something interesting, we can pick it back up.
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Old 01-03-2014, 11:42 PM   #240
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* invalidate perpetuated myths about fishless cycling
What particular myths are in your crosshairs? It seems like this thread has a lot of good information, but lacks goal-oriented research. I think you would benefit from selective only a few (for now) to use as a jumping off point for your research, adding new questions as they become relevant or as you become satisfied with your previous ones.
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