Justfrozen's Fishless Cycle Log

[waterdrop]

Hi Mr Bliss. I agree completely with your comments about nitrification in the substrate. I believe though that articles in Applied and Environmental Microbiology have provided evidence that the primary nitrite oxidizers are Nitrospira spp. and not Nitrobacter. I believe there are also beginning to be articles in the journals related to waste water treatment plant operation that are also moving away from Nitrobacter to Nitrospira.

The photophobic aspect is interesting. I had thought that this aspect of these autotrophic species was not well known but our sources as hobbyists is generally poor, so I'd be prepared to believe this is possibly correct.

~~waterdrop~~

 

[Mr Bliss]

Hey thanks for reminding me about that, waterdrop! This is probably the billionth time I read about this, but Nitrospira just doesn't want to stay in my brain.

Nitrosomonas is definitely photophobic. It was one of the bacteria we had to isolate during an advanced microbiology course back in the day, and protecting the samples from light was essential. Apparently one of the ammonia-oxidizing enzymes can't stand (blue) light. A couple of old abstracts about it:

http/www.ncbi.nlm.nih.gov/pmc/articles/PMC245697/
http/www.ncbi.nlm.nih.gov/pubmed/3922353

edit: This article (PDF document) deals with this too, including Nitrobacter. No idea how Nitrospira fits into all this though, although it seems reasonable to assume they use the same or very similar enzymes and are inhibited by the same things.

 

[waterdrop]

Oh good, I'm glad we're on the same page about that!

Wow! Thanks for the addition of new articles to read! I've perused the PDF and it looks great. This is the kind of reading I like to take on the bus. Obviously the photophobic characteristic is quite real and I wonder if this might have real implications for clear filter boxes versus darker ones?

~~waterdrop~~

 

[JustFrozen]

Day 17
Ammonia = 0.0
Nitrite = 1.0

With all your advice I'm holding off on the substrate - I need more time and money at the moment anyway.

I'm going to re-dose for the first time tonight. The only problem being that I still don't have a determined amount of ammonia to add. I may just add only 1ml this time, see the results, and increase if necessary.

The nitrite seems to be sticking even though ammonia is dropping. I find that surprising because I thought there was more of a direct relationship between the two. Could this mean I have a reasonable amount of the 2nd type of bacteria?

 

[waterdrop]

Not sure what you are picturing in your mind. Your A-Bac colony seems to have grown and is now working on processing down the large ammonia dose, thus producing a larger amount of nitrite(NO2.) There are a small number of N-Bacs there, steadily processing some NO2 into NO3 but now that the A-Bacs are dumping in more NO2, that amount has surpassed what the N-Bacs can process and the "leftover" is not dropping back below 1.0ppm.

This is exactly what we'd expect to see as it gets later in the first phase and is ramping up for the second phase of fishless cycling. We see signs that ammonia is dropping faster and that the slow rise of the excess nitrite(NO2) is heading upward toward the "nitrite spike" of the second phase. The N-Bacs can be growing at roughly the same slow rate as the A-Bacs but still appear to be fewer in number because the molecules they have to process (NO2) are 2.7 times greater than the food (NH3) that the A-Bacs have to process.

At any rate, your fishless cycle looks to be on track!
~~waterdrop~~

 

[JustFrozen]

I've got another question, now that I'm apparently in the nitrite spike phase.

In the fishless cycle article stickied at the top of the forum it says:

NOTE FOR API TEST KIT USERS: When you add the drops, if they immediately turn purple in the bottom of the tube, your nitrites are off the chart high. You do not need to shake the tube and wait 5 minutes. If you do, the color will turn green as the nitrites are so high that there isn't a color to measure them with.

I've noticed recently that the drops most definitely turn deep purple at the bottom of the tube; however, I still shake and wait five minutes, and the color is far from green. It's still a purplish hue, and it still looks like it's on the chart. I think the above statement in the article is a little misleading in one way or another. Am I wrong in assuming I still have a measurable level of nitrite?

 

[waterdrop]

What's most common is for the drops to turn a deep purple directly as they go into a tube of test water that has a very high nitrite content. If your peak nitrite level is still 1.0 then you are not even seeing the purple as dark and shiny as it gets yet. That "Note" clause that RDD put in there was because a *few* users where encountering an even more rare and even more confusing observation that sometimes when the nitrite was "extremely" high, the drops would take on a weird shiny blue-green look right when the settled in the tube after being dripped in. I've observed this. Usually when the tube is then shaken, the resulting color is still very dark purple, but very occasionally the shaken color is a weird grayish green sort of thing. RDD was just confirming that when you more or less know you're probably still in the nitrite spike phase (often lasts for more than a week or longer) and see these dark shiny purple or blue-green drops form immediately, there's no need to wait through the 5min test, it just means you are "off the chart."

Other than that period of "off the chart" however, I always recommend only judging a color test right at the 5 minute mark (not before, not later) and always using the same lighting setup and to always attempt to judge "hue" and not contrast (although it certainly can be acknowledged that that can be hard sometimes.)

~~waterdrop~~

 

[JustFrozen]

I think I understand what you mean. It's not necessarily that the drops turning purple equals off-the-chart, it's that it's definitely not near 0, which in the nitrite spike phase, is all that matters. Therefore there's no need to wait the 5 minutes, other than recording the actual amount, such as when using them in logs such as this.

Correct?

 

[waterdrop]

Well, I'm like you and always did like having a good record in my log, just so I could understand better if nothing else. I'd continue to wait out the 5 minutes and take a reading -until- they reach 5.0 (that's top of the chart, right?) and stay there for a couple days. After that point, if the shiny sinking initial drops look the same as these 2 days then it seems fine to just write "5+" in your log and rinse the testtube without waiting in my opinion.

Later on, when the end of the nitrite spike comes, it is usually abrupt, taking place in a day or two in my experience watching cases here.

~~waterdrop~~

 

[JustFrozen]

Day 22
Ammonia = 0.0
Nitrite = 0.5
Nitrate = 7.5

My tank is consistently cycling 4ppm ammonia within 24 hours. The nitrite has been dropping and is almost completely gone. Also nitrate is climbing but still seems very low. I'm not sure if my tank is starting to struggle or if I'm on the verge of the final phase. I could believe it only because of the slightly mature media that was added early on, but considering the nitrate is so low, it seems I've got a long way to go still in processing Nitrite

I think I need to re-read the guide again

 

[OldMan47]

It looks like you are making nice progress Justfrozen. If it feels like things are slowing, check the pH of the water. A low pH will slow things. The thing to be aware of with the nitrate test is that it will give low readings if you don't mix the chemicals well enough. I usually double the shaking and mixing times they suggest.The problem is that things settle in the reagent bottles and until they are mixed back in properly you are really not getting a good test.

 

[waterdrop]

Sounding good to me too. Hours it takes to drop ammonia and nitrite is a much more definate sign ultimately than any sort of feedback from the unpredictable nitrate. Its nice with nitrate corroborates the other two, but not something I worry about.

~~waterdrop~~

 

[JustFrozen]

Day 23

I'm not sure what happened tonight. I must have been distracted. The reading for ammonia came out at 6.0

I thought to myself, that can't be right... and I retested it twice to be sure. Sure enough, I got 0.0 for both additional readings. I'm not sure what I did wrong before.

So that might be a good tip - if you get a reading that is definitely strange, consider re-testing instead of doing something to counter-act it

 

[JustFrozen]

Day 24
Ammonia = 0.0
Nitrite = 5+

Notes: I realize now I've been making a mistake. I thought the nitrite was falling, but it is clearly off the chart. It has been for the past few days. I've gone back and updated those days to reflect this.

On one hand I'm further back in the cycle process than I thought. On the hand, I now have the experience of this to fall back on, and now I know what to look for.

Had I listened to RDD and WD more , this probably wouldn't have happened. I guess I need to pay better attention

Also, on a side note, I've got a theory as to what happened with that strange ammonia test result I've got. As I've mentioned before I often leave the test results in the tubes overnight and then clean them before use the next day. I also use one of the tubes to measure the ammonia. That night the ammonia tube was the only one empty after everything was done. The next day I either assumed it had been cleaned and just used it straight away, or didn't clean it enough before using it for the ammonia test. Considering how concentrated that stuff is, I bet it would only take a fraction of a drop to wildly skew the results.

At least at the end of the day, the cycle is now doing what I expect of it. Hopefully I can manage the rest of this without a hitch

 

[OldMan47]

When I do a test, I dump it down the sink right away after reading the results. Then I rinse the test tube several times with tap water and put the wet test tube upside down in a small bowl to drip dry. By the time I want to do another test, I have a nice clean dry tube with no residue in it. I did go out on E-bay and buy a dozen of the test tubes because I am clumsy and had broken a few. It was getting to the point where if I wanted to test for ammonia, I had to rinse out the nitrite tube that I had just used. That is no way to run a test program. Now that I have plenty of replacements, I seldom seem to break them any more. It is almost a Murphy's law, you only break things when there is no spare to use.